CHAPTER ONE

1.1 INTRODUCTION

The Student Industrial Work Experience Scheme (SIWES), also known as Industrial Training is
a compulsory Skills Training Program designed to expose and prepare students of Nigerian
Universities, Polytechnics, Colleges of Education, Colleges of Technology and Colleges of
Agriculture, for the industrial work situation they’re likely to meet after graduation.

The scheme also affords students the opportunity of familiarizing and exposing themselves to the
needed experience in handling equipment and machinery that are usually not available in their
institution.

Before the establishment of the scheme, there was a growing concern among industrialists, that
graduates of institutions of higher learning lacked adequate practical background studies
preparatory for employment in industries. Thus, employers were of the opinion that the
theoretical education in higher institutions wasn’t responsive to the needs of the employers of
labour.

SIWES introduction, initiation and design was done by the Industrial Training Fund (I.T.F) in
1993 to acquaint students with the skills of handling employer’s equipment and machinery.

The Industrial Training Fund (I.T.F) solely funded the scheme during its formative years.
However, due to financial constraints, the fund withdrew from the Scheme in 1978.

The Federal Government, noting the significance of the skills training handed the management of
the scheme to both the National Universities Commission (N.U.C) and the National Board for
Technical Education (N.B.T.E) in 1979.

The management and implementation of the scheme was however reverted to the I.T.F by the
Federal Government in November, 1984 and the administration was effectively taken over by the
Industrial Training Fund in July 1985, with the funding solely borne by the Federal Government.

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1.2 HISTORY OF SIWES

SIWES was established in 1973 under the military administration of Gen. Yakubu Gowon,
former head of state of the federal republic of Nigeria. This program is supported and sponsored
by the industrial training fund (ITF), with approval of the National Board of Technical Education
(NBTE), and acceptance of the federal government. SIWES is a vital venture of almost all
professional and technical training programs all over the world. This program was designed to
expose students’ participation in the practical working experience during the attachment.
Students are assigned to specific duties which are to be supervised by the industry- based
supervisors to enhance the skills of the participating students.

The scheme started in 1974 with 48 students from 11 institutions of higher learning participating.
By 1978, the scope of participation in the scheme has increased to about 5000 students from 32
participating institutions. The Industrial Training Fund withdrew from the management of the
scheme in 1979 wing to problems of logistics and the increased financial burden associated with the
rapid expansion of SIWES. Consequently, the Federal government funded the scheme through the
National Universities Commission (NUC) and the National Board for Technical Education (NBTE)
who managed SIWES for five years (1979-1984). The supervising agencies (NUC and NBTE)
operated the scheme in conjunction with their respective institutions during the training. Later, in
November 1984, the federal government revert the management and implementation of the scheme
to ITF. In July 1985, it was taken over by the Industrial Training Fund (ITF) while the funding was
solely borne by the federal government.

1.3 AIMS OF SIWES

SIWES is strategized for skill acquisition. It is in fact designed to prepare and expose students of
universities, polytechnics and colleges of education to the real-life work situation they would be
engaged in after graduation. Therefore, SIWES is a key factor required to inject and help keep alive
industrialization and economic development in the nation through the introduction and practical
teaching of scientific and technological skills to students.

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1.4 OBJECTIVES OF SIWES
 To complement the tertiary institutions in the provision of adequately trained manpower.
 To complement the tertiary institutions in the provision of adequately trained manpower.
 To complement the institutions in improving indigenous technology.
 To expose SIWES students to technicalities and methods of handling equipment
andmachines which, are not usually available in their institutions.
 To prepare students for work situations they may likely meet after graduation.
 To enable students put their theoretical knowledge into practice.
 To enable the students on return to their institutions, strike a balance between their
practical experience and their theoretical knowledge.
 To give room to employers of labour to make their own input into the nation's
educational process.

1.5 BRIEF HISTORY OF RAYSCAN LABORATORY AND MEDICAL SERVICES

Rayscan Laboratory and Medical Services is a diagnostic centre that came into existence in the
year 2015. They started with scanning in a small room with just three staffs and they're doing so
well with so many achievements because of the accuracy of their results.

The quality of what they gave made them to be having lots of patients and a lot of patients were
referred to the centre. As time goes on the laboratory services and other investigations came into
existence which they're perfectly running several tests and investigations with more than 25
staffs from different departments in a bigger place and also they currently have 2 branches at
Gyadi-Gyadi court road, and Audu Bako way, Kano State.

1.6 SERVICES RENDERED AT RAYSCAN LABORATORY AND MEDICAL

SERVICES
 Routine laboratory tests to cater for the Health needs of the public through the various
departments they have which include: medical microbiology department, Haematology
department, virology department, serology department, and clinical chemistry unit.
 Medical laboratory investigations
 X-ray services
 Ultrasound scans
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  Colonoscopy
 Endoscopy
 And many more.

1.7 THE DIFFERENT DEPARTMENTS IN THE ORGANIZATION

Reception

Specimen collection unit

Laboratory Department

X-ray department

Doppler unit

Ultrasound Scan Unit

CHAPTER TWO

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Rayscan Laboratory and Medical Services aims to cater for the welfare of people as regards to
their health by carrying out routine laboratory tests through the various departments present in
the facility. Experiences gained throughout the period of attachment are summarized according
to the unit sent to for attachment.

2.1. SPECIMEN COLLECTION UNIT

This department specializes in Specimen collections which include blood Specimen, stool
Specimen, urine specimen, sputum specimen, swab specimen such as wound swab, throat swab
and High Vaginal Swab (HVS).

2.1.1. Collection of Blood Specimen

Collecting blood specimen is a very sensitive process therefore; blood must be collected with
adequate care to ensure safety of the technologist and reliability of results. Protective gloves should
be worn when collecting blood specimen. Lancets, needles, and syringes must be discarded safely
to avoid accidents and injury. Blood specimen can be collected using two methods; capillary and
vein.

2.1.2. Capillary Blood Collection Technique

Capillary blood collection technique is used when the volume of blood required to run a test is
small. Capillary blood can be used to make thick and thin blood film; it can be used to measure
Packed Cell Volume (PCV) and also used for blood group test. Capillary blood can be collected
from the finger of a child or adult and from the heel of an infant up to one year old. Though
capillary blood collection is an easy technique of blood collection, it has some disadvantages as
follows: it can be used for only few tests, greater possibility of specimen error, difficulty to
obtaining sufficient blood, platelets cannot be estimated in blood films made from capillary blood.

Materials;

Lancet

Cotton wool, and a pair of hand gloves.

Procedure

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  Puncture site was cleaned using 70% ethanol and allowed to air-dry.
 Using lancet or pricker pricks the sterilized area of the finger.
 Pressure was applied to the finger pricked to enhance blood flow. The first drop of blood
was wiped off with cotton wool and the next drop was used to run the test to avoid body
juice.
 When sufficient blood has been collected, a piece of cotton wool swab was pressed over the
pricked surface until bleeding stopped.

2.1.3. Venous Blood Collection Technique

Venous blood is used when more than 100µl of whole blood is required to run a test, when serum
from clotted blood specimen is needed, when plasma from un-clotted blood specimen is needed and
when more than one test is to be conducted. Venous blood can be used for antibody or antigen test.
Venous blood is preferable to capillary blood for reason previously mentioned, particularly when
the patient is an adult and several tests are required.

Apparatus

Syringe and needle,

Tourniquet

Cotton wool and a pair of hand gloves.

Procedure;

Before collection, the blood specimen container was labeled with the patient’s name, time and date
of collection.

 A soft tubing tourniquet was applied to the upper arm of the patient.
 Using the index finger, a suitable vein (a large straight vein that does not roll and with a
direction that can be felt) was selected.
 The puncture site was cleansed using 70% ethanol and allowed to dry after which the patient
was instructed to tighten his or her fist.

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  With the thumb of the left hand holding down the puncture site, a puncture was made with
the level of the needle directed upward in the line of the vein.
 The plunger of the needle was steadily withdrawn at the speed it took the vein to be filled.
 When sufficient blood was collected, the patient was instructed to release his or her fist.
 The needle was removed gently and the puncture site was immediately pressed with a piece
of dry cotton wool until bleeding stopped.
 The needle was removed from the syringe and the blood sample container which can be an
EDTA (ethylene Diamine tetra acetic acid) bottle or a plain test tube was filled with the
required volume of blood.
 The needle and syringe were safely discarded to avoid injury and infection.

2.1.4. Collection of High Vaginal Swab (HVS) Specimen;

A vaginal swab test involves taking a sample of vaginal secretions with a device that looks like a
cotton bud. The swab, with secretions attached, is then placed in a sterile swab container.

Apparatus

Speculum

Swab stick, and a pair of hand gloves.

Procedure;

 A sterile vaginal speculum moistened with warm water was inserted into the vaginal canal.
 Once inserted, the speculum was rotated back 90 degrees (so that the handle is facing
upwards).The speculum blades were opened until an optimal view of the cervix was
achieved.
 The locking nut was tightened to fix the blades in position.
 The swab stick was taken out of the container and inserted into the Vaginal opening; it was
rotated at 360 degrees to get the sample.
 The swab stick was removed gently and placed in its container.

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2.1.5. Collection of Sputum Specimen;

The patient was given a clean, dry, wide-necked, leak-proof, screw cap container and requested to
breathe in and exhale several times and then cough deeply to produce the sputum specimen.

NOTE: The specimen must not contain saliva, mucus or nasal secretion and is best collected in the
morning, soon after the patient wakes up and before any mouth wash is used.

2.1.6. Collection of Stool Specimen

The patient was given a clean, dry, disinfectant-free wide-necked container and the following
instructions were given to the patient on how to obtain the specimen:

 Transfer a portion of the stool specimen especially that which contains mucus, pus or blood
in to a clean, dry, leak proof container.
 After collection of the specimen from the patient, it was labeled and sent to the
microbiology bench for processing.

2.1.7. Collection of Urine Specimen

 The patient was given a sterile, dry, wide-necked, leak proof container and about 10-20ml of
the specimen was requested. The following instructions were given to the patient on how to
collect the urine.
 Female patients wash their hands, clean the area around the urethral opening with clean
water, allow to air dry and collect the urine with the labia apart. Allow the first few drops of
urine to pass before collecting the specimen mid-stream. Also, male patients were asked to
wash their hands before collecting the urine specimen (middle of the urine flow).
 The container was labeled with the patient’s name, reference number, date and time of
collection.
It was then delivered as soon as possible to the laboratory with the request form for processing.

2.1.8. Collection of Swab Specimen (Wound swab, Ear swab, Throat swab)

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  A sterile swab stick (see fig. 1 in appendix) was used to wipe the infected area which can be
wound in any part of the body, ear or throat making sure that the specimen was picked.
 The swab stick was returned back into its container and closed tightly to avoid
contamination.
 It was sent to the laboratory for further examination.

2.2. MEDICAL MICROBIOLOGY BENCH

Medical microbiology bench is an important bench in any laboratory. It is concerned with the
diagnosis and treatment of infectious diseases. In addition, it also studies various clinical
applications of microbes for the improvement of health. There are four kinds of microorganisms
that cause infectious disease: bacteria, fungi, parasites and viruses. The test conducted in medical
microbiology bench deals with the use of instruments such as incubator, autoclave, microscope,
petri dishes and the most important of which is culture media.

2.2.1 The Principles of Culture Media

To study and identify microorganisms, they have to be cultivated and isolated in pure culture. Most
bacteria can be cultured in artificial media if the media meet the nutritional and physical growth
requirements of the bacteria. These media must contain water and sources of nitrogen, carbon,
mineral salts and vitamins. Therefore, the principle governing the use of culture media is based on
the fact that microbes can grow in a medium where there is sufficient nutrients and favourable
environmental conditions.

2.2.2 Types of Media

For a culture media to be successful in growing microorganisms, it must contain all essential
nutrients, ions and moisture, maintain the correct pH, osmotic pressure, and neutralize any toxic
material produced by the microorganism. Culture media are of different forms including solid
media, semi-solid media, liquid media depending on the amount of the solidifying agent (agar)
added. The main types of culture media used in routine microbiological culture techniques include:

1. Basic media: These are simple media such as nutrient agar and nutrient broth which support
the growth of most microorganisms that do not need special nutritional requirements. These
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 media are used in the preparation of enriched media and enrichment media and also for sub-
culturing of pathogens from differential and selective media prior to performing
biochemical test.
2. Enriched media: These media support the growth of microorganisms with specific growth
requirements such as Haemophilus influenza, Neisseria gonorrhoaeand some Streptococcus
species. Basic media can be enriched with whole or lysed blood, serum, special extracts or
nutrients to support the growth of bacteria that cannot grow on the basic media. Examples of
these media include; blood agar and chocolate agar.
3. Selective media: These are solid media, which contain substances that slow down or inhibit
the growth of microorganisms other than those for which they have been devised. Example
of selective media include telluride medium for Diphtheria spp, Deoxycholate Citrate Agar
(DCA) and Salmonella Shigella Agar (SSA) for Salmonella and Shigella groups.
4. Enrichment media: These are liquid media and are similar in function to selective media.
An example of enrichment media is Selenite F broth used for the isolation of Salmonella
species.
5. Differential media: These media contain substances or indicators that differentiate one
organism from another. Examples of this media include MacConkey agar, which
differentiates lactose fermenting from non-lactose fermenting bacteria, blood agar, which
differentiates haemolytic bacteria from non haemolytic bacteria and Cysteine Lactose
Electrolyte Deficiency Agar (CLED) use to differentiate between lactose fermenter and
mom lactose fermenting microbes present in urine Specimen.
6. Transport media: These are media meant for transportation of clinical specimens that
contain delicate microorganisms in instances where there is to be a delay in their delivery to
the laboratory for processing. They are generally semi-solid e.g. Amies transport medium
for Neisseria gonorrhoae and Stuart’s medium or Cary Blair medium.
2.2.3 Preparation of Media

Culture media are prepared according to the manufacturer’s instructions as indicated on the
container label of the powdered media. Examples are;

MacConkey agar 52g/litre

Deoxycholate Citrate agar 70.52g/litre

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 Nutrient agar 28g/litre

Saborouddextrous agar 56g/litre

CLED. 36.25g/litre

For instance, if 80mls of nutrient agar is to be prepared, it is done using the following
procedures;

28 grams---------1000ml

Then x grams -------80ml

Then, = 28g x 80ml=1000 x x grams = 2.24 gram

Apparatus

Weighing balance, autoclave, burnsen flame, measuring cylinder, conical flask, filter paper, spatula
and a pair of gloves.

Procedure

 The weight of the paper was determined and confirmed to be negligible before weighing the
powdered media using the value obtained from the calculation above using weighing
balance.
 The weighed powdered media was then disolved in 80mls of water and autoclaved at 121ᵒC
for 15 minutes.
 It was then allowed to cool to 45ᵒC after which it was poured into dishes with the lid of the
dishes left half closed to solidify.
 After solidification, it was inverted and stored in incubator to dry.(See fig. 2 in appendix
showing media preparation).

2.2.4. STAINING TECHNIQUES

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Identification of bacteria involves a standardized process that first uses a Gram stain or other stain
to partially characterize the organism. This information may fully identify a particular bacterial
genus or even species but further testing is essential to identify the organism. Common staining
techniques used in medical microbiology are the gram stain and Zheilnelson acid fast stain.

Gram Staining Technique

The Gram stain technique was developed in 1884 by the Danish physician Christian Gram and is
the most widely employed staining method in the study of bacteria. It is an example of differential
staining because it is used to distinguish organisms based on how they react to the different kinds of
stains used in the technique. Gram stain divides bacteria into two classes namely gram negative and
gram positive. It is used for the staining of bacterial cells only. It reveals the shape (morphology)
and arrangement of the cells present in the preparation.

Principle of Gram Stain.

The difference in Grams reaction between bacteria species is attributed to the difference in
permeability of the cell wall of Gram positive and Gram-negative organisms during the staining
process. Following staining with crystal violet and iodine and on treatment with acetone, the dye-
iodine complex is easily removed from the more permeable cell wall of Gram-negative bacteria but
not from the less permeable cell wall of Gram-positive bacteria. A counter stain is applied to give
Gram-negative bacteria a pinkish or red colour. While Gram positive bacteria retain the colour of
the primary stain and thus stain purple.

Procedure

 A drop of normal saline was applied on a clean grease-free slide.

 With the aid of a sterile wire loop, a colony of the test organism was picked and
emulsified in the normal saline.
 The smear was allowed to air-dry completely and heat-fixed by passing it 2-3 times on a
Bunsen burner flame.
 Placing it on a staining rack, the slide was stained with crystal violet stain for 1 minute
after which it was washed off running water.

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  Lugol’s iodine was applied to the smear and was allowed to act for 1 minute and washed
off with running water.
 It was decolourized with 70% ethanol and washed immediately with running water.
 The smear was counter stained with safranin and was allowed to act for 1 minute after
which it was washed with running water and allowed to air dry.
 A drop of immersion oil was placed on the smear and it was examined using the x100
objective.
Results;

Gram positive bacteria stain dark purple while gram negative bacteria stain pinkish or red.

TABLE 1: Examples of gram positive and gram negative bacteria

Gram positive Bacteria Gram negative bacteria

Staphylococcus spp. Escherichia coli

Clostridium spp. Shigella sp.

Streptococcus spp. Salmonella spp.

Corynebacterium Neisseria spp.

Spp.

Klebsiella spp. Vibrio cholerae

Lactobacillus spp. Proteus spp.

Listeria spp. Serratiamarcescens

2.2.5. ANTIMICROBIAL SUSCEPTIBILITY TESTING

In the treatment and control of infectious diseases, especially when caused by pathogens that are
drug resistant, sensitivity test is used to select effective antimicrobial drugs that are used to treat

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such infections. The test is conducted to determine the antibiotics that can treat an infections disease
and those cannot.

Principle

Laboratory antimicrobial susceptibility test can be performed using the disc diffusion techniques. A
disc of blotting paper impregnated with a known volume of antimicrobial agent is placed on a
susceptibility-testing agar uniformly inoculated with test organism. The antimicrobial agent diffuses
from the disc into the medium and the growth of the organism is inhibited at some distance from the
disc. However, resistant strains have smaller zone of inhibition or grow to the edge of the disc.

Apparatus

Wire loop, agar plate, sensitivity disc (either gram positive or negative).

Procedure

 An agar plate that is free from contamination was passed over flame 2-3 times to
sterilize.
 A young colony of the isolated microorganism was picked from the primary culture and
uniformly streaked over the surface of the sterile agar.
 Using a sterile forceps, the antimicrobial sensitivity disc (see fig. 4 in appendix) was
picked from its container and placed on the inoculated agar plates, ensuring that each
disc was pressed down lightly to touch the surface of the agar.
 The plate was incubated aerobically at 35-37ᵒC for 24hour
The size of the zones of inhibition of each antimicrobial agent can be interpreted using the
interpretative chart, reporting the result as resistant, intermediate or susceptible.

Resistant: When there is no zone of inhibition around the disc.

Intermediate: A cloudy plaque or zone at some distance from the disc indicates that not all species
of bacteria were cleared.

Sensitive: A clear circular hollow around the disc indicates that the antibiotics have completely
inhibited the growth of the microorganisms. Sensitive results and intermediate results were reported
as 6+, 5+, 4+, 3+, 2+, and a + depending on the size of the zone of inhibition.
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2.2.6. STOOL MICROSCOPY, CULTURE AND SENSITIVITY (M/C/S)

The stool m/c/s is a test that detects and identifies bacteria that cause infections of the lower
digestive tract. The test distinguishes between the types of bacteria that cause disease (pathogenic)
and the types that are normally found in the digestive tract (normal flora). The test helps to
determine if pathogenic bacteria are the cause of a person's gastrointestinal symptoms
(gastroenteritis).The stool will be checked for colour, consistency, amount, shape, odour, and the
presence of mucus.

Macroscopic Examination

This involves the physical examination of the stool sample using parameters such as:

 Colour appearance: either brown, black, green, or yellowish brown.

 Consistency: either formed, semi-formed or unformed.
 Constituents: that is, the presence of blood (at the surface or mixed), mucus, pus,
live and/or segments of tapeworm.

Microscopic Examination

Stool sample is examined microscopically using wet preparation

Apparatus

A clean grease-free slide, applicator stick, stool specimen container, microscope

Reagents

Normal saline or Lugol’s iodine, applicator stick, glass slide, cover slip and microscope.

Procedure

 A drop of iodine or normal saline was dispensed on a clean grease-free glass slide.

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  Using a sterile wire loop or applicator stick, a pea-size of the stool specimen was picked and
emulsified in the saline or iodine solution.
 A cover slip was mounted avoiding air bubbles and over floating.
 The slide was examined using the 10x objective lens and then 40x objective lens to confirm
any parasite seen. (See fig. 5 in appendix showing stool microscopy).

Procedure for stool culture and sensitivity

 Xylose Lysine Deoxycholate Agar (XLD) was prepared following manufactures

instruction.
 The media was sterilized using the autoclave and allowed to cool down to 45°c.
 It was dispensed into sterile petri dish and allowed to solidify.
 Using a sterile wire loop or applicator stick, the stool sample was picked aseptically and
streaked on the surface of the media.
 It was turned upside down and incubated at 37°c for 24 hours. (See fig. 6 in appendix
showing bacteria growth from stool culture).
If microbial growth is present after incubation, sensitivity test is conducted using the following
procedures;

 Nutrient agar was prepared following manufactures instruction and autoclaved.

 It was dispensed into a sterile petri dish and left to cool.
 Using a wire loop, an isolate of the microorganism was picked and spread on the surface
of the medium.
 A pair of forceps was used to aseptically pick the sensitivity disc and it was placed over
the surface of the medium. It was pressed gently to enable it stick to the surface.
 It was incubated at 37°c for 24 hours.

2.2.7. URINE MICROSCOPY, CULTURE AND SENSITIVITY (URINE M/C/S)

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Urine is one of the body’s waste products. It is produced in the kidneys and collected in the bladder
until a person urinates. Normally, the urine does not contain significant numbers of any
microorganism. However, if bacteria or yeast are introduced into the urinary tract, they can multiply
and cause a urinary tract infection, called a UTI. Therefore, urine m/c/s is used to diagnose a
urinary tract infection (UTI).

Macroscopic examination

The physical appearance of the urine samples was observed and recorded using parameters such as;

Colour appearance: either amber, pale amber or yellow.

Nature: either clear or cloudy.

Odour: Pungent smell, foul smell and so on.

TABLE 2: Appearance of urine specimen in diseased condition

APPEARANCE POSSIBLE CAUSE

Cloudy usually with unpleasant smell and and
Bacterial urinary tract infection.
white blood cells

Red and cloudy due to red blood cells. Urinary Schistosomiasis, bacterial infections.

Brown and cloudy due to haemoglobin Conditions that cause intravascular

haemolysis

Yellow or green-brown due to bilirubin Acute viral hepatitis, obstructive jaundice

Yellow-orange due to urobilinogen Haemolysis, hepatocellular jaundice

Microscopy
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Urine specimen is usually examined microscopically using wet preparation to detect pus cells, yeast
cells, motile parasite, ova of, white blood cells and bacteria in freshly collected urine specimen.

Apparatus

Clean greese free slide, cover slip, wire loop, microscope.

Procedure

 Using a sterile wire loop, a drop of the urine specimen was fetched and dispensed on a
clean grease-free microscopic slide.
 A cover slip was mounted on the drop of urine avoiding air bubbles and over-floating.
 The slide was examined microscopically using the 10x and 40x objectives.

Culture and sensitivity

Culture of urine specimen was always performed whenever the urine contains bacteria, casts, or
nitrates. Cysteine Lactose Electrolyte Deficient agar is widely used by most laboratories to isolate
urinary pathogens because it gives consistent results and support the growth of both gram positive
and gram negative organisms. Sabouraud Dextrose Agar (SDA) is also used in urine culture to
isolate fungi such as Candia albicans.

Apparatus
Sterile wire loop,

Culture plate, containing prepared media.

Bunsen flame.

Procedure

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  The freshly collected clean-catch sample was mixed by rotating the container gently
without shaking.
 Using a sterile wire loop, a loopful of the specimen was picked and streaked on the
surface of CLED agar.
 The plate was incubated aerobically at 37ᵒC for 24 hours. (See fig. 7 in appendix
showing the process of urine culture).
 Nutrient agar was prepared and colonies isolated from the culture were sub-cultured in
it.
 Using a pair of sterile forcep, an antibiotic sensitivity disc was placed on the inoculated
plate and it was incubated at 37°c for 24 hours.
 After incubation, the sensitivities of the bacteria to the various antibiotic suspensions
was read and recorded.

2.2.8. HIGH VAGINAL SWAB MICROSCOPY, CULTURE AND SENSITIVITY (HVS

M/C/S)

HVS is a test conducted to help diagnose vaginal infections that don’t affect the urinary tract. The
causes of vaginitis can include bacteria such as Streptococcus spp, Clostridium spp, E.coli,
Staphylococcus spp, etc. Yeast infections caused by Candida albicans or viruses.

Microscopy

HVS microscopy is usually conducted using wet mount technique

Apparatus

High vaginal swab stick, normal saline, microscope, glass slid and cover slip.

Procedure

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  Speculum was used to open the vagina by rotating it.
 A swab was made using the swab stick by inserting it into the patient’s vagina and
rotating it gently while it was still in the vagina.
 After swabbing, the swab stick was immediately returned to the pack to preserve the
specimen.
 A drop of normal saline was placed on a clean grease-free slide.
 The swab was emulsified in the drop of normal saline.
 A cover slip was mounted on the smear avoiding air bubbles and over-floating.
 The slide was examined microscopically using the 10x and 40x objective lenses.
Appearance epithelial cells, pus cells and yeast cells were recorded.

Culture and sensitivity

HVS specimen can be cultured on blood agar, MacConkey agar, chocolate agar or SDA

Apparatus

Sterile wire loop, culture plate containing prepared media, Bunsen burner flame

Procedure

 The swab stick containing the specimen was inoculated in the media and incubated at
37ᵒC for 24hours.
 After incubation, the plate was observed for growth or appearance of colonies on the
surface of the media.
 When bacterial growth was confirmed, nutrient agar was prepared and the bacteria
isolate was sub-cultured in it.
 An antibiotic sensitivity disc which is either gram positive or negative was placed on the
inoculated media and incubated at 37ᵒC for 24hours. After incubation, the sensitivities of
the bacterial pathogens to the antibiotics were determined.

2.2.9. SPUTUM MICROSCOPY.

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It is conducted to help diagnose tuberculosis (TB) and infections caused by other Mycobacterium
species, which are known as acid-fast bacilli (AFB). It is also used to diagnose other
microorganisms that cause lung infections such as Streptococcus spp.

Macroscopic examination

Here, the physical appearance of the sputum specimen was observed and described:

Purulent: green looking musty pus.

Mucopurulent: Green looking with pus and mucus.

Mucoid: Mostly mucus

Mucosalivary: Mucus with small amount of saliva.

Microscopy

 A smear was made from the sputum specimen on a glass slide and stained by the gram
staining techniques.
 A drop of immersion oil was placed on the smear which was mounted on the stage of a
microscope and examined for pus cells and predominant bacteria e.g. Streptococcus
pneumonia, Haemophilusinfluenzae, and Staphylococcus aureus.
 Another smear was made on a glass slide and was stained using the Ziehl-Neelson
techniques to detect Acid-Fast Bacilli (AFB) in the case of blood tint sputum.

2.3. WOUND SWAB MICROSCOPY, CULTURE AND SENSITIVITY

Wound swab m/c/s is done to diagnose pathogenic microorganisms associated with open wounds.
These organisms are opportunistic in nature. They gain entrance into the body through open
wounds and cause other infections from there. Pathogenic organisms associated with wounds
include; Streptococcus spp, Klebsiellaspp, Proteus spp, E.coli etc.

Microscopy

21
It is done using wet preparation

Procedure

 A sterile swab stick was used to swab the infected area.

 The swab stick was returned back to its container to avoid contamination.
 A drop of normal saline was placed on a clean grease-free slide.
 The swab was emulsified in the drop of normal saline.
 A cover slip was mounted on the smear avoiding air bubbles and over-floating.
 The slide was examined microscopically using the 10x and 40x objective lenses.

Culture and Sensitivity

 The specimen collected using the swab stick was inoculated into already prepared media
and incubated at 37°C for 24 hours.
 When bacterial growth was confirmed, it was sub-cultured into freshly prepared nutrient
agar.
 Forcep was used to pick an antibiotic sensitivity disc which was placed over the surface
of the medium.
 It was incubated at 37°C for 24 hours. (See fig. 8 in appendix showing incubator).
After incubation, the sensitivities of the microbes was determined.

2.4. HEMATOLOGY BENCH

The Hematology Laboratory performs routine hematology testing, limited coagulation, and various
body fluid testing and analysis. It is concerned with the study of the cause, prognosis, treatment,
and prevention of diseases related to blood.

2.4.1. BLOOD GROUP TEST

ABO and Rhesus blood grouping systems are clinically important. Blood donors and recipients
must be correctly ABO grouped because transfusing incompatible blood may result in death of a
patient.

Apparatus

22
Blood lancet, cotton wool, white tile, and slide.

Reagents

Anti-A, Anti-B, and Anti-D monoclonal antibodies

Principle

The principle behind blood grouping is based on antigen-antibody reaction leading to agglutination.
Blood sample usually contain antigens that are tested for using the monoclonal antibodies present in
the test reagent and when combined, it agglutinates.

Procedure

 The blood sample was collected by capillary puncture of the thumb using blood lancet.
 A clean grease-free tile was divided into three sections and a drop of blood was placed
on each section.
 A drop of each antiserum was placed in each of the divided portion.
 It was mixed to get a homogenous mixture.
 The tile was rocked by tilting gently from side to side while taking note of agglutination.
Results;

Agglutination with A and D antisera means the blood group is A positive denoted A+

Agglutination with only A antisera means the blood group is A negative denoted A-

Agglutination with B and D antisera means the blood group is B positive denoted by B+

Agglutination with only B antisera means the blood group is B negative denoted B-

Agglutination with A, B, and D antisera means the blood group is AB positive denoted AB+

Agglutination with A and B antisera means the blood group is AB negative denoted AB-

Agglutination with only D antisera means the blood group is O positive denoted O+

No agglutination at all, means the blood group is O negative denoted O-

Application of blood grouping

23
Blood grouping is used for compatibility testing between blood donor and recipient before
transfusion. It can also be applied in marriage counseling in order to prevent the occurrence of
certain disease condition such as sickle cell anaemia.

2.4.2. MALARIA PARASITE TEST

Malaria parasite test is carried out to detect the presence of Plasmodium sp. in the blood of a
patient. Five species of plasmodium cause malaria in man which are; Plasmodium falciparum,
Plasmodium ovale,Plasmodiummalariae, Plasmodium vivax and Plasmodium knowensi. Of all
these species, Plasmodium falciparum is common in Nigeria and other tropical and sub-tropical
regions.

Malaria parasite is transmitted by the bite of an infected female Anopheles mosquito. Sporozoites
contained in the saliva are inoculated in the human blood when the mosquito takes a blood meal.
Malaria parasite can be examined microscopically using thick and thin blood film preparations. The
test is also conducted using a Malaria Rapid Diagnostic Test ( RDT) kit specific to P.falciparum.

Procedure for making thick blood films

 The lobe of a finger was cleaned with the aid of swab moistened with 70% alcohol and
allowed to air-dry.
 Using a sterile lancet, the finger was pricked and squeezed gently to obtain a large drop
of blood.
 The first flow of blood was wiped off to reduce tissue juice and next drop was used to
make the film.
 The drop of blood was placed in the center of a sterile grease-free clean glass slide.
 Using a spreader, the drop of blood was spread to make a thick film.
Staining procedure for thick blood films

 The dried thick film was placed on a staining rack.

 It was covered with field stain A and allowed to act for 2 seconds and rinsed off with
water.

24
  It was stained with field stain B and allowed to act for 2 seconds after which it was
rinsed off with water.
 It was allowed to air dry.
 A drop of immersion oil was placed on the film and it was focused using the 100x
objective lens.

Making thin blood films

 A drop of venous blood was placed on the edge of a clean grease-free slide.
 Using a smooth edge spreader or another glass slide with smooth edge, the film was
spread to make a thin film by drawing the glass forward to touch the blood film and
allowing the blood film to spread through the edge.
 Holding the spreader at an angle of about 45ᵒC, the blood was spread by pushing the
spreader forward and releasing it to make a thin film,( see fig. 10 in appendix) and
allowed to air-dry.
Staining procedure for thin blood films

 The dried thin film was placed on a staining rack.

 The slide was covered with Leishmans stain and allowed to act for 2minutes (fixing
period).
 It was double diluted with distilled water and allowed to act for 8minutes.
 Excess stain was washed off using clean water and allowed to air-dry.
 A drop of immersion oil was placed on the body and tail part of the film and it was
focused under the microscope using the 100x objective lens.
Reporting Malaria Parasites

About 100 microscopic fields were observed and the number of parasites seen was reported as
follows:

+ means 1-10 parasites per 100 thick film field.

++ means 11-100 parasites per 100 thick film field

+++ means 1-10 parasites per single thick thin film field
25
++++ means more than 10 parasites per single thick film field

MALARIA PARASITE TEST USING RDT KIT

Malaria RDT kit is used to rapidly detect the presence of P. falciparum antigens present in a
patient's blood.

Principle

The membrane slip of RDT kit is coated with mouse monoclonal antibodies specific to
P.falciparum antigen. When the sample is placed in the sample well, it reacts with mouse antibodies
and moves chromatographically along the membrane to the test region producing a visible line
which indicates a positive result while absence of such line indicates a negative result.

Apparatus

RDT kit, lancet, cotton wool swab.

Reagent

Malaria test assay diluent.

Procedure

 An alcohol swab was used to sterilize the patient's finger to be pricked.

 A sterile lancet was used to prick the finger, the first drop of blood was wiped off and
the second drop of blood was placed in the sample well of the RDT kit.
 Two drops of the assay diluent was dispensed in the assay diluent well and two drops
was dispensed into the sample well.
 The result was read after 10 minutes.

2.4.3. ERYTHROCYTE SEDIMENTATION RATE (ESR)

Erythrocyte sedimentation rate (ESR) is a test that indirectly measures the degree of inflammation
present in the body. The test actually measures the rate of fall (sedimentation) of erythrocytes (red
26
blood cells) in a sample of blood that has been placed into a tall, thin, vertical tube. Results are
reported as the millimeters of clear fluid (plasma) that are present at the top portion of the tube after
one hour.The ESR is not diagnostic; it is a non-specific test that may be elevated in a number of
these different conditions. It provides general information about the presence or absence of an
inflammatory condition.

Principle

When citrated blood in a vertically positioned Westergreen pipette is left undisturbed, red cells
aggregate, stack together to form a rouleaux and sediment through the plasma, the ESR is the rate at
which this sedimentation occurs in 1hour as indicated by the column of the clear plasma above the
red cells measured in millimetre.

Apparatus

Westergreen ESR pipette, Westergrene ESR stand, timer, Pasteur pipette.

Procedure

 About 2-5ml of blood was collected from a patient through venous blood collection.
 The blood was transferred into a capped filling tube containing sodium Citrate as
anticoagulant and was shaker vigorously to get a homogenous mixture.
 Westergreen pipette was inserted into the capped filling tube, the mixture was drawn to
the zero mark of the Westergreen pipette avoiding air bubbles.
 The Westergreen pipette was placed on the Westergreen ESR stand and left undisturbed
for 1hour.
 After 1hour, the level at which the plasma meet the blood was read and recorded in mm.
Reference range of ESR value (normal)

Male 3-5mmfall/hour

Female4-7mmfall/hour

27
 Infant 0-2mmfall/hour

Clinical significance of ESR test

High ESR values are obtained during the period of menstruation and pregnancy. Since there are
several factors that can affect cell sedimentation rate, slightly and moderately raised values cannot
be significant. ESR can be used to diagnose a wide range of diseases.

Causes of significantly high ESR values may include Tuberculosis, HIV/AIDS, Acute viral
hepatitis, Bacterial endocarditis, heart disease, kidney disease, cancer, rheumatic fever, etc.

Causes of lower ESR values may include: polycythemia, allergic state, sickle cell anaemia, dengue
haemorrhagic fever, leucocytosis, etc

2.4.4. PACKED CELL VOLUME TEST (PCV)

Packed cell volume is used to screen for anaemia when it is not possible to measure haemoglobin
accurately. PCV is used in the investigation of burns, dengue haemorrhagic fever and polycythemia.
Principle

The packed cell volume is the portion of whole blood occupied by red cells expressed as a ratio
(liter/liter). Anticoagulated blood in a glass capillary of specific length, bore size and wall thickness
is centrifuged at 10,000rpm for 5 minutes to obtain a constant packing of red cells. A small amount
of plasma remains trapped between the packed red cells. The PCV is red from the scale of
haematocrit reader.

Apparatus

Heparinized capillary tube for PCV, plasticine,haematocrit centrifuge, microhaematocrit reader.

Procedure

 Blood specimen collected through the vein was transferred into an EDTA tube and
mixed gently.

28
  Heparinized capillary was inserted into the blood sample and the blood moves by
capillary action into the capillary tube.
 One end of the capillary tube was sealed using plastacin.
 The capillary tube was inserted into one of the numbered slots of the haematocrit
centrifuge with the sealed end against the rim gasket.
 The specimen was centrifuged at 10000 rpm for 5 minutes.
 After centrifugation, the heparinized capillary was removed from the haematocrit
centrifuge.
 The PCV reading was taken using the haematocrit reader. (See fig. 13 showing PCV
result reading using haematocrit reader).
Reference range of PCV value

Male 40-45%
Female 30-37%
Children 50-70%
Women at menopause 40-45%

Clinical Significance of PCV

In a similar way to haemoglobin levels, PCV values vary with age, gender and altitude. Also,
ranges may vary in different population and laboratories. A decreased PCV indicates anaemia.
Further testing may be done to determine the exact cause of the anaemia. Conditions that can result
in low PCV include: bleeding, nutritional vitamin, iron or other mineral deficiencies, inflammatory
conditions such as rheumatoid arthritis, kidney diseases, cirrhosis, haemolysis, bone marrow
disorder and pregnancy. The most common cause of increased PCV is dehydration.

2.5. CLINICAL CHEMISTRY BENCH

Clinical chemistry department is a department that is generally concerned with analysis of bodily
fluids for diagnostic and therapeutic purposes.
29
2.5.1. URINALYSIS TEST

Urinalysis is a series of tests on urine. Doctors use it to check for signs of common conditions or
diseases such as kidney, liver or heart diseases, diabetes etc. This can be determined by by checking
the amount of protein, glucose, bilirubin, urobilinogen, ketones, blood, nitrites, pH and ascorbic
acid present in the urine.

Apparatus

Medi combi-9 container (see fig.14 in appendix) with urinalysis strips, specimen container, hand
gloves and urinalysis chart.

Procedure

 The lid of the urine specimen container was unscrewed.

 A strip was aseptically removed and immersed in the urine for few seconds.
 Excess urine was removed from the strip by tapping the strip be gently.
 After 30-60seconds, the reaction was carefully read by comparing it with the chart on
the container label avoiding contamination
Result

The result of urinalysis test can be determined using the following parameters:

Blood:

The presence of blood in urine indicates parasitemia, due to Schistosomiasis,

Trichomonasvaginalis, bacterial infections and pregnancy.

Urobilinogen:

This indicates abnormal haemolysis due to hepatocellular disease, sickle cell, Hepatitis and malaria.
Obstructive jaundice and liver diseases

Protein:

This can be found in the urine of patients with urinary schistosomiasis, kidney diseases, urinary
tract infections and pregnancy.

30
Nitrite:

This indicates urinary tract infections caused by nitrate reducing bacteria such as Klebsiellaspp,
E.coli, Proteus spp, Enterobacterspp, Psueudomonas spp.

Ketones:

This can be in the urine of untreated diabetic patients, it can also occur due to starvation,
dehydration, pregnancy and vomiting.

Ascorbic acid:

This indicates high intake of fruits and vitamins, and problems within the glomerular filtrate.

Glucose:

Presence of glucose in urine indicates deficiency in insulin level, glucosuria and other conditions
causing diabetes and candidiasis.

Leukocytes:

This can be found in the urine of patients with urinary tract infection (UTI) and inflammation.

PH:

Abnormal pH range (below or above 5.1-8.0) may be due to drug intake, bacterial infection caused
by Proteus and Klebsiella spp and kidney stones.

Bilirubin:

Presence of bilirubin in the urine indicates liver disease such as hepatitis, liver paranchymal disease,
excessive hemolysis and constipation.

2.5.2. FASTING BLOOD GLUCOSE OR SUGAR TEST (FBG OR FBS)

Fasting blood sugar is one of the known techniques for measuring blood glucose or plasma level. It
is used to screen a patient for diabetes mellitus. Good control of blood glucose may help prevent or

31
delay the development of complications, which may lead to premature disability or death from
blindness, kidney failure and bacterial infections. The test can be conducted using a glucometre or a
spectrophotometer.

Preparation for the test

Patients were instructed to fast for about 8hours before running the test. They are usually adviced to
come for the test in the morning.

FBG using Glucometer:

Glucometer is an instrument used to measure blood glucose levels.FBG test is done using a strip
inserted into the strip port of a glucometer with blood sample in it.

Principle

The test reaction involves glucose in the blood specimen which is oxidized to glucunolactones in
the reactive zone of the strip. During this reaction, hexacyanoferrate III is reduced to
hexacyanoferrate II. A voltage is applied between the two palladium electrodes contained in the
strip. This reaction leads to the hexacyanoferrate II being deoxidized to hexacyanoferrate III with
the release of electrons. This small electric current that is generated is measured by the meter and
converted into a glucose concentration reading.

Apparatus

One touch ultra-2 glucometer, strip, 70% alcohol, cotton wool, sterile lancet and hand gloves.

Procedure

32
  The test strip was inserted into the glucometer and it automatically turn on the
glucometer.
 The finger of the patient was swabbed with alcohol and a sterileancet was used to prick
the finger.
 The blood specimen obtained through capillary puncture was placed on the edge of the
glucometer.
 The result was automatically displayed digitally on the screen of the glucometer and the
value gotten was divided by 18 to get the actual result. (See fig 15 in appendix showing
FBG test).
Approximate glucose reference

Fasting adults: 3.9-5.6mmol/L

Children: 2.4-5.3mmol/L

Fasting Blood Glucose Using Spectrophotometer

A spectrophotometer is an instrument that measures the rate of absorbance of a substance at

different wavelengths. It is also used to conduct both Fasting blood glucose and random blood
glucose.

Principle

A spectrophotometer works based on Beard Lambert's law which states that the rate of absorbance
of light by a sample is directly proportional to the concentration of the sample at a specific
wavelength. A spectrophotometer is made up of two devices; a spectrometer and a photometer.
Spectrometer produced light of different wavelength, while a photometer measures the intensity of
the light produced. The solution sample is placed between the spectrometer and the photometer for
the sample to be analysed.

Apparatus

Centrifuge, spectrophotometer, cuvette, test tubes.

Reagents

33
Standard solution, test reagent.

Procedure

 2mls of Venous blood was collected from a patient and was dispensed into anticoagulant
fluoride oxide tube to avoid clotting after which it was centrifuged.
 The plasma was dispensed into a sterile clean container.
 1000uL of the test reagent was pipetted into 3 test tubes representing blanck, standard
and sample.
 10uL of standard solution was pipetted into the standard test tube.
 10uL of the sample (plasma) was pipetted into the sample test tube and allowed to stand
for 25 minutes.
 The spectrophotometer (see fig. 16 in appendix) was switched on and set at a
wavelength of 505.
 The cuvette was rinsed with distilled water and the blanck solution was dispensed into
the cuvette which was placed in the spectrophotometer to blanckit( calibrating it to
0.00).
 The standard solution was dispensed into the cuvette and placed on the
spectrophotometer and the reading was taken. The cuvette was rinsed with distilled
water.
 The sample was dispensed into the cuvette which was placed in the spectrophotometer
and the reading was taken. The cuvette was also rinsed with distilled water.
 The result was calculated using the formulae
Absorbance of sample ÷ absorbance of standard × 5.55

The result gotten is recorded in mmol/l

2.5.3 RANDOM BLOOD SUGAR OR GLUCOSE (RBS)

34
This is a test conducted to check the amount of blood sugar present in the blood. It is different from
Fasting blood sugar because this test can be done at any time of the day and does not require
fasting. It is also conducted using glucometre and it's strip. It can be used to diagnose diabetes and
other sugar related diseases.

Apparatus

Glucometer

Glucometre test strip

Hand gloves and lancet.

Procedure

 Swab the patient's finger to be pricked with alcohol swab.

 Prick the finger and place a drop of blood on the glucometre test strip already inserted
into the glucometre.
 The result was taken and divided by 18 to get the actual result.
Result

Non diabetic adults Less than 11.1mmol/L

2.5.4. PREGNANCY TEST OR HUMAN CHORIONIC GONADOTROPIN TEST (HCG)

Human chorionic gonadotropin (HCG) is a glycoprotein hormone produced by placental cell soon
after the fertilized ovum is planted in the uterine wall. Laboratory testing is based on the detection
of rapidly rising level of HCG in urine or blood serum or plasma.

Principle

When the specimen is applied to the absorbent pad, the antibody-dye conjugate binds to the HCG
antibody in the serum forming HCG antibody-antigen complexes. These complexes migrate to the
reaction zone where it binds to the HCG forming a coloured band (pink-rose in colour). In a

35
negative test, no coloured band is produced in the patients’ zone. Unbound conjugate binds to the
reagent in the control zone producing a coloured zone indicating a correctly performed test.

Apparatus

Specimen container, HCG test strip

Procedure

 First, early morning urine is recommended because it contain the highest level of HCG.
 The absorbent pad of the strip was dipped in the urine sample for 15seconds.
 The strip removed and placed on a work bench horizontally for migration to occur.
 The result was read after 1minute.
 Alternatively, serum obtained from venous blood can be used in place of urine following
the same procedures.
Result

Appearance of two thick lines in the control and test region of the test strip indicates a positive
result.

Appearance of a thick line only in the control region indicates a negative result.

Appearance of a thick line only in the test region or absence of thick line in both the patient and test
region indicates an invalid test.

2.5.5. FAECAL OCCULT BLOOD TEST (FOB)

Many diseases can cause hidden blood in stool in the early stage of gastrointestinal problems such
as ulcer, colon cancer and so on. The presence of blood in the stool indicates peptic ulcer caused by
a gram negative bacteria Helicobacter pylori.

Principle

The membrane of the faecal occult blood test strip is pre-coated with antibody. The mixture
migrates upward on the membrane chromatographically by capillary action to react with the anti-

36
haemoglobin antibody on the membrane of the coloured line on the test region indicating a positive
result. While its absence indicates a negative result

Procedure

 The patients’ faecal specimen was collected in a clean specimen bottle.

 8-10 drops of the faecal occult blood test buffer was dispensed in a test tube.
 A pea-size of the faecal specimen was added to the FOB buffer and mixed homogenously.
 The faecal occult blood test strip was immersed and allowed to act for 2-3 minutes before
removing it.
 The result was read and recorded.
Result

Appearance of two thick lines in the control and test region of the test strip indicates a positive
result.

Appearance of a thick line only in the control region indicates a negative result.

Appearance of a thick line only in the test region or absence of thick line in both the patient and test
region indicates an invalid test.

2.5.6. HELICOBACTER PYLORI TEST

Helicobacter pylori is a bacterium that causes peptic ulcer in individuals.

Apparatus

H.pylori test casette, test buffer, specimen container and hand gloves.

Procedure

 Faecal sample was collected in a specimen container.

 Pea size of the specimen was picked and dispensed into the test buffer solution.
 The mixture was mixed to get a homogenous mixture.
 Two to three drops of the solution was dispensed into the specimen well.
 Result was read after 10 minutes.
Result
37
Appearance of two thick lines in the control and test region of the test strip indicates a positive
result.

Appearance of a thick line only in the control region indicates a negative result.

Appearance of a thick line only in the test region or absence of thick line in both the patient and test
region indicates an invalid test.

2.6. SEROLOGY BENCH

Serology test deals with antibody-antigen reaction. Virology department is usually concerned with
carrying out test that detects the presence of viruses in individuals. Common viruses tested for are
retrovirus causing HIV and viruses affecting the liver which are hepatitis B and C viruses.

2.6.1. HEPATITIS B SURFACE ANTIGEN TEST (HBSAg)

Hepatitis is a disease that causes damage to the liver and is characterized by the presence of
inflammatory cells in the tissue or organs. Hepatitis is caused by a double stranded DNA virus of
the family hepanaviridae. The virus can be detected by serological test using blood plasma or serum
after centrifugation.

Principle

Hepatitis B surface antigen test depends on the principle of antigen-antibody reaction. The
membrane of the test strip is incorporated with Anti-HBsAg antibody on the test line region of the
strip. During testing, the serum reacts with the particle coated with anti-HBsAg antibody. The
mixture migrates chromatographically by capillary action thereby reacting with the HBsAg
generating a coloured line.

Apparatus

HBsAg test strip, EDTA tube, Pasteur pipette, cotton wool, 70% alcohol, needle and syringe, tubing
tourniquet and hand gloves.

Procedure

38
  2ml of the patient’s blood sample was collected and poured into a clean EDTA tube.
 The blood specimen was centrifuged at 3000rpm for 5minutes to obtain the serum.
 The specimen application zone of the test strip was dipped into the serum and allowed to
stand for 5-10minutes for optimum migration to occur by capillary action.
 The strip was removed and the test result was read and recorded.
Result

Appearance of two thick lines in the control and test region of the test strip indicates a positive
result.

Appearance of a thick line only in the control region indicates a negative result.

Appearance of a thick line only in the test region or absence of thick line in both the patient and test
region indicates an invalid test.

2.6.2 HEPATITIS C VIRUS SERUM/PLASMA TEST

One-step HCV serum/plasma test is a rapid chromatographic immunoassay for the qualitative
detection of antibody specific to Hepatitis C virus (HCV) in serum or plasma. The test strip and
serum have to be at room temperature before carrying out the test.

Principle

The membrane of HCV test strip is coated with HCV antigen on the test line of the strip. During
testing, the mixture migrates chromatographically by capillary action to react with the HCV antigen
on the membrane and generates a coloured line which indicates a positive result.

Procedure

 The strip was removed from the pack and carefully dipped into the serum/plasma obtained
from venous blood avoiding contamination.
 After a few moments the strip was removed and placed horizontally on a clean, dry non-
absorbent surface.
 The result was read after 15seconds when the serum/plasma has migrated through the
control and test region of the strip.
Result
39
Appearance of two thick lines in the control and test region of the test strip indicates a positive
result.

Appearance of a thick line only in the control region indicates a negative result.

Appearance of a thick line only in the test region or absence of thick line in both the patient and test
region indicates an invalid test.

2.6.3 RETROVIRAL SCREENING TEST (RVS)

The test is used to screen the blood of donors and recipients before transfusion and also to
determine the HIV status of an individual which can help in reducing the pandemic nature of the
disease. HIV cause progressive impairment of the body’s immune system leading to increased
susceptibility to diseases and tumors and the fatal condition called AIDS (Acquired Immune
Deficiency Syndrome). HIV can be present in virginal cervical secretion and blood which are the
main routes through which the virus is transmitted.

Apparatus

EDTA tube, syringe and needle, tubing tourniquet, 70% alcohol, cotton wool, Pasteur pipette,
centrifuge, hand gloves, HIV-1 and HIV-2 determine strip.

Principle

The HIV-1 and HIV-2 determine strip test is an immune chromatographic test for the detection of
antibodies to HIV-1 and 2 in a patient’s serum. As the serum moves through the conjugated pad, it
reconstitutes and mixes with selenium colloid antigen conjugate on the strip. This mixture migrates
through the solid phase to the immobilized recombinant antigen colloid and synthetic peptide at the
patient’s window site. If the antibodies to HIV-1 and 2 are present, it binds to the antigen selenium
colloid and to the antigen of the patient forming a red band at the patient’s window site. If the
antibodies to HIV-1 and 2 are absent, the unbound selenium colloid antigen migrates and passes the
patient’s window site to the control band where it forms a red coloured band indicating the validity
of the test.

Procedure

40
  2ml of blood sample was collected from the patient via the vein and transferred into a clean
EDTA tube.
 The specimen was centrifuged at 3000rpm for 5minutes to obtain the serum.
 Using Pasteur pipette, the serum was drawn and applied to the specimen or specimen zone
of the determine test strip.
 The result was read after 15minutes. (See fig. 18 in appendix for RVS test kits).

Result

Appearance of two thick lines in the control and test region of the test strip indicates a positive
result.

Appearance of a thick line only in the control region indicates a negative result.

Appearance of a thick line only in the test region or absence of thick line in both the patient and test
region indicates an invalid test

2.6.4 WIDAL TEST

This is an agglutination test usually carried out to detect antibodies specific to the O and H antigens
of Salmonella in the serum of a patient with enteric fever. It is a screening test that uses
agglutination reaction to diagnose typhoid fever and other Salmonella infections. Therefore, the
result of the test can be confirmed by the quantitative method of testing (culturing). The H antigen
is the flagellate antigen and it is responsible for headache and other related symptoms. The O
antigen is the somatic antigen and it is responsible for stomach ache and related symptoms.

Apparatus

A clean sterile grease-free tile, Pasteur pipette, blood collection equipment, centrifuge

Reagents

Febrile antigen test kits (widal kits) containing concentrated smooth antigen suspension of both
Salmonella typhi O with their respective paratyphi (A,B,C) a positive or negative control.

Principle
41
Widal test is based on agglutination reaction that test for the presence of antibodies against
Salmonella organism that causes Typhoid fever. The antibodies are serum products which in
response to exposure to salmonella organism will agglutinate the bacterial suspension which carries
homologous antigens.

Procedure

 2-5ml of the patients’ blood was collected through venous blood collection and poured into
an EDTA tube.
 The tube was balanced by another tube containing an equal proportion of any fluid placed in
opposite direction in the rotor of a centrifuge.
 The centrifuge was switched on and the blood was spun at 3000rpm for 2minutes to separate
the serum from the blood.
 A small volume of the serum was drawn from the tube using the Pasteur pipette and
dispensed in eight different positions on a white tile.
 Equal drops of Salmonella Typhi O and H antigens with their respective paratyphi (A, B, C)
were applied to the corresponding drops of serum.
 Using the edge of a clean grease-free slide or mixer, the drops of blood and serum were
mixed properly avoiding contamination.
 The tile was rocked for 60seconds while observing for agglutination reaction. (See fig. 19 in
appendix for widal test).
Result

Agglutination that occurs within the first 15seconds and becomes more prominent after rocking was
given the ratio 1:160.

Agglutinations occurring within the first 30seconds and become less prominent compared to the
first instance after rocking were given the ratio 1:80.

The same thing applied to 1:40 at 45seconds and 1:20 at 60seconds with the least amount of
agglutinates.

42
NOTE: Variations were observed as some agglutination might start late but become more
prominent after rocking. In such cases they were also given the ratio 1:80. The ratios 1:320, 1:160,
and 1:80 are significant results while 1:40 and 1:20 are insignificant results.

2.6.5 VENERAL DISEASE RESEARCH LABORATORY (VDRL) TEST

Syphilis is a chronic sexually transmitted infection caused by the spirochete bacteria,

Treponemapallidum, resulting in the formation of lesions (spores) through the body.

Apparatus

VDRL test strip, centrifuge, blood collection equipment (needle and syringe, EDTA tube, cotton
wool, spirit, tubing tourniquet)

Procedure

 2-5ml of the blood was centrifuged to obtain the serum.

 Using a sterile Pasteur pipette, the serum was collected from the centrifuge tube and applied
on the specimen region of the test strip placed horizontally on the workbench.
 Migration occurs through capillary action from the specimen region through the control and
the test or patient’s band.
Result

Appearance of two thick lines in the control and test region of the test strip indicates a positive
result.

Appearance of a thick line only in the control region indicates a negative result.

Appearance of a thick line only in the test region or absence of thick line in both the patient and test
region indicates an invalid test.

CHAPTER THREE

3.1 EXPERIENCE GAINED

43
  Knowledge on the operation and working principles of some machines and equipment used
in the laboratory. Examples of these equipments are incubator, centrifuge, microhematocrit
centrifuge, etc.
 Identification of pathogens in clinical specimens and cultures and presentation of results of
laboratory tests.
 Requisition and analysis of specimen based on marked symptoms presented by a patient.
 Interpretation of results of various tests conducted in the laboratory. These tests are strip
tests such as Hepatitis B and C test, HIV test, syphilis test. Other test arewidal test, PCV etc.
 Ability to collaborate effectively with others in carrying out complex tasks.

3.2 PROBLEMS ENCOUNTERED

 Lack of financial support from the organization.
 Refusal of accepting microbiology students in medical laboratories.
 Problem of getting a place for industrial attachment.
 Little knowledge on the working principles of some machines found in the laboratory.

3.3 SOLUTIONS PROFFERED

The challenges were overcome in the following ways:

 Asking questions about how the machines work.

 More organizations should be enlightened on the need of accepting students on industrial
training.
 The law restricting microbiology students from working in the laboratory should be revised
so as to avoid enable microbiologist work in medical laboratory.

CHAPTER FOUR

4.1 CONCLUSION

44
The Students Industrial Work Experience Scheme (SIWES) affords students the opportunity of
familiarizing and exposing themselves to the needed experience in handling equipment and
machines that are usually not available in their institution. Therefore, during the period of my
industrial attachment at Rayscan Laboratory and Medical Services, I acquired practical knowledge
on some of the theoretical work done in school.

During the period of my attachment, I partook in routine activities taking place in the various
benches of the laboratory which are: collection and registration of specimen, preparation of culture
media, culture and sensitivity testing, identification of bacterial pathogens (using the Gram staining
techniques, and biochemical tests), malaria parasite test, microscopic examination of stool, urine,
high vaginal swab, sputum samples, urinalysis test, widal test, retroviral screening test, and hepatitis
test among others.

Through these activities, my understanding of the activities done theoretically was greatly improved
as I was exposed to it practically. The attachment has brought about new knowledge and skills,
good interaction with people, ability to work well with others (team work) and better understanding
of laboratory work practices.

4.2 RECOMMENDATIONS

The Student’s Industrial Work Experience Scheme (SIWES) has exposed students to sophisticated
machines and equipment, professional work methods e.t.c. It also trains students to prepare for
future employment and challenges that they might face after schooling.

Therefore, it is based on these reasons stated above that I recommend the continuity of the students
industrial attachment program as it helps to bridge the gap between theory and practice. I also
recommend that more organizations be enlightened on the need for participation so as to ease the
training of students.

REFERENCES

45
Burtis, C. A. ;Ashwood, E. R.; Bruns, D. E. (2006). Tietz Textbook of Clinical Chemistry. (4th
ed.) Saunders. pp. 2448. ISBN 978-0-7216-0189-2

Cheesbrough, M. M. (2005): District Laboratory practice in Tropical countries. (2nd ed.) Part
1.pp 195-295

Fritz, H. K.; Kurt A. B.; Johannes, E. D.; Rolf M. Z.: (2005). Kayser Medical Microbiology
(10th ed.) Thieme Stuttgart. New York. pp 147, 164 ISBN 3-13-131991-7

Madigan, M. T.; Martinko, J. M.; (2006). Brock Biology of Microorganisms (11th ed.). Pearson.
ISBN 978-0-13-196893-6

Ochei, J. and Kolhatker, A. (2005): Medical Laboratory Science Theory and Practical. pp 254-
720

Ryan, K. J.; Ray, C. G.; (2004).Sherris Medical Microbiology: an introduction to infectious

diseases (4th ed.) McGraw-Hill. ISBN 978-0-8385-8529

https://siwesbeginner.com/siwes-introduction

APPENDICES

46
 Figure 1: Swab sticks
Figure 2: Antimicrobial sensitivity disc

Fig 3: Stool culture showing bacterial growth Fig 4; Thick blood film

Source: learning.unonbi.com

Fig 5; Incubator Fig 6; RVS Kit

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Fig 7: Medicombi 9

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