A branch of science that deals with the

study of body fluids.

A branch of science that deals with the

determination of the type and characteristics
of blood, blood testing, and examination of
blood stain, semen, saliva and other body
fluids that may or may not be involved with
DNA typing.

Blood can be used as circumstantial or corroborative evidence in identifying

the perpetrator of a crime. It can also be used to prove disputed percentage
as paternity test, determine the cause of death, or the length of time at
which the victim survived an attack.

1
As circumstance or corroborative evidence against
or in favor of the perpetrator of the crime.

2
As evidence in case of disputed percentage

As evidence in the determination of the cause of

3 death and the length of time the victim survived the
attack.
4 Determination of the direction of escape of the
victim or the assailant

5 Determination of the origin of the flow of blood.

As evidence in the determination of the

6 approximate time the crime was committed.
 Is the circulating tissue of the body. It is
referred to as highly complex mixture of cells,
enzymes, proteins and inorganic substances. It is
the red fluid of the blood vessels.
Blood is opaque. On the treatment with either,
water or other reagents becomes transparent lake
color. It is finally alkaline. Normally pH is 7.35
– 7.45.

(45%) Formed elements or the solid materials consisting

chiefly of cells namely:

Red Blood Cells or RBC (Erythrocytes)

contains hemoglobin and carry oxygen to various cells in the

body.

White Blood Cells or WBC (Leukocytes)

are masses of nucleated protoplasm. Help fight infection.

Blood Platelets (Thromobocytes)

cells that are produced by the bone marrow and are necessary
for proper clothing of blood. Normally responsible for the
reaction of blood clot.
The fluid or liquid portion where the cells are suspended.
It is principally composed of:

Water – 90% Solid – 10%

The yellowish fluid of blood in which numerous

blood corpuscles are suspended. A straw-yellow
liquid formed when blood to which oxalate has
been added to prevent clotting is allowed to
strand.

A straw-yellow liquid
formed when clotted blood
is allowed to stand for
some time and the clot
contracts.
 Where has to be searched for

Collection, preservation, packing and transportation of

specimen suspected to contain blood.

Blood offers little resistance to decomposition.

It undergoes a rapid charge in its character with
the passage of time as process of clotting and
drying commences almost immediately on exposure
to air.
Sodium fluoride maybe added to blood to preserve
it for a week at room temperature or indefinitely
in a refrigerator. Between 40 – 50 degrees
centigrade is the ideal preserving temperature for
blood and other perishable specimens.
Collection of blood stains should be done as soon as possible; mere washing
of garments/clotting removes the blood.

COLLECT FROM

Victims of Crimes of
violence

Parent and child in case

of disputed parentage.

DRIED BLOOD OR BLOOD

Bulky objects like
Smooth surface like walls,
blackboard, linoleum sheets,
finished floors, table tops
doors, window frames
Glazed surface like glass,
tiles, automobiles. Clothing

Hard surface like axe, hammer, Blood absorbed by

knives, stones, crowbars the soil
 PRELIMINARY TEST
Determine whether the stain contains blood
or another substance. Determines whether
visible stains do or do not contain blood.
It is used to demonstrate the presence of
blood.

CONFIRMATORY TEST

Determines whether bloodstain really contains

blood. Test that positively identifies blood.

PRECIPITIN TEST

Determines whether blood is a human or non-human origin, and if

non-human. The specific animal family from which it originated.

BLOOD GROUPING TEST

Determines whether blood is a human or non-human origin, and if

non-human. The specific animal family from which it originated.

FOLLOWING ARE PRELIMINARY TEST FOR BLOOD:

1 Benzidine Test or Benzidine Color Test

Phenolphthalein Test (also known as Kastle-

2 Meyer Test)
X
Guaiacum Test (Van Deen Test,
3 Day’s or Schonbein’s Test)

4
Leucomalachite Green Test

5
Luminol Test
 BENZIDINE TEST
An extremely sensitive test that can be applied to
minute stain. For many years the most commonly used
preliminary test for blood.

The Benzidine test never fails to detect blood

even when very old, decomposed stain will sort of
contamination is examined.

The positive result is only indicative that

the blood maybe present.

REAGENT POSITIVE RESULTS

Benzidine solution (small Intense blue color
amount of powdered benzidine produced immediately
dissolved in glacial acetic
acid) and 3% solution of
hydrogen peroxide.
LIMITATION
Benzidine test is not a specific test for blood. Positive results may be
obtained from substance as sputum, pus, nasal secretion, plant justice,
formalin, clay, gun. The reaction is weaker and produce fain coloration.

PHENOLPHTALEIN

An alternative test to benzidine test. It

can detect blood in a dilution of 1:80, 000,
000 parts.

A positive result with this test is highly

indicative of blood.

REAGENT
Phenolphthalein solution (1-2 grams’
phenolphthalein to 100 mL of 25% KOH in water
added with one gram zinc powder heated until
colorless) and 3% solution of hydrogen peroxide.

POSITIVE RESULTS LIMITATION

Rose color develops or deep Test is also given by copper salts,
pink color or permanganate potatoes and horseradish.
color.
 GUAIACUM TEST

A fairly delicate test showing the

presence of fresh blood in a solution
of 1:50,000 dilution.

It may not react to very old stains.

REAGENT POSITIVE RESULTS

Fresh tincture of guaiac Beautiful blue color that
resin (Few lumps of this to appears immediately.
95% alcohol, then filter)
and 3% of hydrogen peroxide
or few drops of turpentine.
LIMITATION
The test also reacts with saliva, pus, bile, milk, rust, iron salts, cheese,
gluten, potatoes, perspiration and other oxidizing substances.

LEUCOMALACHITE

Not as sensitive as the benzidine test

REAGENT
Leucomalachite Green Solution (1 gram
leucomalachite green dissolved in 48 mL, glacial
acetic acid and diluted to 250mL water) and 3%
hydrogen peroxide.

POSITIVE RESULTS
Malachite green or bluish green
 PEROXIDASE
Enzyme that accelerates the oxidation of several
classes of organic compounds by peroxide.

HEMOGLOBIN
The red coloring matter of the red blood cells of the blood.

LUMINOUS TEST
An important presumptive identification test for blood. The reaction
of luminol with blood result in the production of light rather than
color. By spraying luminol reagent onto a suspect item, large areas
can be quickly screened for the presence of bloodstains.
The sprayed object must be located in a darkened area while being
viewed for the emission of light (Luminescence).

Luminol test is extremely sensitive test. It is capable of detecting

bloodstains diluted up to 10,000x. Luminol is known to destroy many
important blood factors necessary for the forensic characterization
of blood, so its use should be limited only to seeking out blood
invisible to the naked eye.

Microscopic Test
THE THREE
THE THREE CONFIRMATORY
CONFIRMATORY TEST
TEST
FOR BLOOD
FOR BLOOD (OR
(OR THE
THE THREE
THREE TEST
TEST Microchemical Test
TO DETERMINE
TO DETERMINE IFIF
STAIN IS
STAIN IS
REALLY BLOOD)
REALLY BLOOD)
Spectroscope Test
Useful for the demonstration and
mensuration of blood corpuscles for making
the distinction between mammalian, avian,
piscine, and reptilian blood and for the
investigation of menstrual lochia and
nasal charges.
In short, it differentiates mammalian,
avian, piscine and reptilian blood.

MAMMALIAN RED BLOOD CELLS

Circular, biconcave disc without nucleus

birds, fish and reptiles red blood cells
larger, oval and nucleated Amphibians-
animal living on land breeding in water.
Red blood cells are larger than mammals,
oval and nucleated.

1 TEICHMANN HEMIN REACTION/TEICHMAN TEST/HAEMIN CRYSTAL

TEST
2 HAEMOCHROMOGEN CRYSTAL TEST OR TAKAYAMA TEST

3 ACETONE-HAEMIN TEST
 HAEMOCHROMOGEN CRYSTAL TEST OR TAKAYAMA TEST

One of the two popular microchemical test. A

delicate test for the presence of hemoglobin.

REAGENT
Takeyama reagent (3 cc. of 10% NaOH, 33cc.
pyridine, 3 cc. of saturated glucose solution and
diluted with 7mL of water.

POSITIVE RESULTS
Large rhombic crystals of a salmon pink color
arranged in clusters, sheaves and other forms that
appear within to 6 minutes when viewed under the
low power objectives. To hasten result heat maybe
applied.

The almost delicate and reliable test for the

determination of the presence of blood in
both old recent stains.
This is performed by means of an optical
instrument known as SPECTROSCOPE.

PRECIPIN TEST
Is the standard test used to determine whether the stain/blood
is of human or animal origin.

REAGENT Precipitin/antiserum

POSITIVE RESULTS
A white cloudy line or ring or band at the contact
points of the fluid that appears immediately or
within one or two minutes

LIMITATION
The precipitin reacts not only with blood proteins
but also with other body proteins as those as
saliva, semen, mucus and other body fluids.
 Blood grouping test of fresh blood (Direct Technique
Method) Using the A-B-O System

THE FOUR BLOOD GROUPS

GROUP O GROUP A GROUP B GROUP AB

AGGLUTINOGEN OR ANTIGEN
These are characteristic chemical
structures or “principles” that the
found on the surface of each red blood
cells which stimulates the production of
agglutinins or antibodies.
There are two different agglutinogens or
antigens classified as Agglutinogen A or
Antigen A and Agglutinogen B or Antigen
B.

ANTIBODY OR AGGLUTININ
These are properties or “principles”
contained in the serum which cause
agglutination or clumping together of
the red blood cells. They are
antitoxic substances within the body
which reacts when confronted with a
specific antigen to protect the
system.
There are two different agglutinins
classified as Anti-A and Anti-B.
Agglutinins are demonstrable in about
50% of newly born infants.

We have the four groups because of the presence of absence of two

antigens A and B in the RBC and two agglutinins Anti-A and Anti-B in
the serum.
Blood Antigen/Agglutinogen Antibodies/Agglutinies
Group Present in the RBC present in the serum
A A Anti-B
B B Anti-A
AB A and B No A and No B or None
O No A and No B or None Anti-A and Anti-B
(+) Means agglutination or clumping of RBC
(-) Means absence of agglutination or no clumping of RBC
 INHERITANCE OF BLOOD GROUPS
Knowledge of the gas of genetics will make it easier to
understand the principle involved in the inheritance of blood
groups.
The inheritance of blood groups is predetermined by the
presence and absence of two facts or GENES called Gene A and
Gene B. Before we understand the inheritance of blood groups
following are definition of items:
GENES
Any of the complex chemical units in the
chromosomes by which hereditary
characters are transmitted. Responsible
for the transmission of hereditary
characteristics. They occur in pair.
There are two genes or factors called Gene
A and Gene B.

These are found in the chromosomes. Since chromosomes go in pair, each

of which carries or fails to carry one of these genes. An individual’s
called genotypes, where O represents the absence in the chromosomes of
either the A or B gene.

CHROMOSOME
S any of the microscopic rod-shape bodies
bearing genes responsible for the
transmission of hereditary
characteristics

PHENOTYPES
term used to denote the expression
of the inherited characteristic as
found in the individual. Actually,
the blood groups.

GENOTYPE
S
are paired genes. It’s either
homozygous or heterozygous
 IMPORTANCE OF BLOOD GROUP DATA
Questions of illegitimacy and relationships in may cause
maybe solved by means of the blood groups as determined by
the agglutinogens A, B, M and N.

Determination of whether a man accused of fathering a child out of

1 wedlock could or could not be its parent.
Determination of whether a child born of a married woman could
2 or could not have been fathered by her legal spouse.
Determination of whether a child could or could not belong to a
3 given set of parents in the case of accidental interchange of
infants in a hospital.

4 Determination of whether a child who has been lost and later

recovered after a long interval could or could not belong to a
given set of parents.

DNA DEOXYRIBONUCLEIC ACID

A chemical substance found in all cells of living organism whose
composition have been passed from parents to offspring. It is called as
the genetic or hereditary material. A person’s DNA is the SAME in every
cell.
Chemically, it is an acid and is composed of
three sub-units, namely:
1. The phosphate group
2. a deoxyribose sugar, and
3. one of the four bases –

Adenine (A)
Thymine (T)
Cytosine (C)
Guanine (G)
DISCOVERY
In 1953, DNA – the so-called building block
or genetic blueprint of life was first
characterized by scientist Francis H. Crick
and James Watson as the material that make
up the genetic code of all organisms.

Crick and Watson were completely unaware of the

potential impact their discovery would have on
forensic science in particular and the criminal
justice system as a whole. Resting on the established
natural truth that no two individuals have the same
DNA blueprint, with the exception of identical twins.

DNA typing or “profilin” as it is now called was

CONCEIVED.
 In 1984 DNA analysis was first recognized as having
application to Forensic Science by the British Molecular
Biologist Alec Jeffreys and thereafter directly used in
a rap and murder case to exonerate and innocent suspect
and to convict the guilty accused.

ALEC JEFFREYS
(1950 – PRESENT)

HOW DNA ANALYSIS IS DONE

DNA Typing is done by first carefully extracting
the DNA from the evidentiary samples.
The DNA is then analyzed to give a particular
pattern.
The patterns are compared with that of a
known individual to determine a match.

SOURCES OF BIOLOGICAL EVIDENCE

BLOOD SEMEN SALIVA URINE HAIR

GENERAL CHARACTERISTICS OF GENOMIC DNA

Each person has a unique DNA profile (except identical
twins because they have the same DNA profile)

Each person’s DNA is the same in every cell (DNA from skin cells
will match from blood cells)

An individual’s DNA profile remains the same throughout life

Half of your DNA comes from your mother and

half from your father
 DNA ANALYSIS

DNA contains “detectable” patterns

unique to each individual

DNA is a robust molecule and is stable

DNA can be isolated from a wide range of biological

samples

Source of is the same in all sources (blood, semen,

sputum, skin etc.)

Precise patterns can now be detected even with very small samples (a
blood sot, single hair follicle, lip-prints on a glass, physical
fingerprints, saliva, skin on a cigarette butt, etc.)

LINE-UP OF CASES WHERE DNA ANALYSIS CAN BE

OF HELP:
SEXUAL ASSAULT HIT AND RUN
CASES LIKE RAPE

MURDER EXTORTION AND BRIBERY

HOMICIDE

ROBBERY PATERNITY CASES

STOLEN OR TRAFFICKED CHILDREN

IDENTIFICATION OF PREVENTION OF ILLEGAL

REMAINS IN MASS ADOPTION OR KIDNAPPING
DISASTER
FIGHT AGAINST HUMAN TRAFFICKING

ESPECIALLY FOR KIDS

 POSSIBLE LOCATION OF
EVIDENCE SOURCE OF DNA
DNA ON THE EVIDENCE
Baseball bat or similar
Handle, end Sweat, skin, blood, tissue
weapon
Hat, bandanna, or mask Inside Sweat, hair, dandruff
Eyeglasses Nose or ear pieces, lens Sweat, skin
Mucus, blood, sweat, semen,
Facial tissue, cotton swab Surface area
earwax
Dirty laundry Surface area Blood, sweat, semen
Toothpick Tips Saliva
Used Cigarette Cigarette butt Saliva
Stamp or envelope Licked area Saliva
Tape or ligature Inside/outside surface Skin, sweat
Bottle, can or glass Sides, mouthpiece Saliva, sweat
Used condom Inside/outside surface Semen, vaginal or rectal cells
Blanket, pillow, sheet Surface area Sweat, hair, semen, urine, saliva
“Through and through
Outside surface Blood, tissue
bullet”
Bite mark Person’s skin or clothing Saliva
Blood, sweat, tissue
Fingernail, partial fingernail Scrapings

GUIDELINES FOR COLLECTING AND SUBMITTING

DNA EVIDENCE
Documenting, collecting, packaging and preserving DNA
Evidence from crime scene.

If DNA evidence is not properly documented, collected, packaged and preserved,

it will not meet the legal and scientific requirements for admissibility in a
court of law.

1 If DNA evidence is not properly documented, its origin can be

questioned.
2 If it is not properly collected, biological activity can be lost.

3 If it is not properly packaged, contamination can occur.

If it is not properly preserved, decomposition and

4 deterioration can occur
 REQUESTS EVIDENCE EXAMINATIONS
All requests for evidence examination should be in writing
addressed to the Director, thru the Deputy Director for
Forensic Investigation Service,
DNA Analysis Laboratory, Forensic Chemistry Division, NBI, Taft
Avenue, Manila and contain the following information:
The requesting and submitting contact person’s name,
1 agency, address and telephone number.
Description of the nature and the basic facts concerning the
2 case (brief history of the case)
The name(s) of and descriptive data about the individuals(s) involved
3 (subject, suspect, victim) and the agency-assigned case
identification number.
4 A list of the evidence being submitted herewith (enclosed) or under
separate cover
5 State what types of examinations are requested

6 State where the laboratory report should be sent.

Chain of Custody of Evidence.

7

PACKAGING AND TRANSPORTING EVIDENCE

1 Take precautions to preserve the evidence
2 Place porous evidence in individual protective covering such as paper
envelopes.

3 Stabilize the evidence to avoid movement or friction during

transport
Wrap and seal each item of evidence separately to avoid
4
contamination.

5 Place the evidence in a clean, dry and previously

unused inner container.

6 Seal the inner container with tamper-evident

tape.

7 Affix EVIDENCE and appropriate BIOHAZARD labels to

the inner and outer container.

8 Affix the evidence examination request and all case

information between the inner and outer container.

9 Place sealed inner contained in a clean, dry and previously unused

outer container with clean packaging material

10 Completely seal the outer container so that opening of the

container would be evident.
 Address the outer container as follows:

FORENSIC CHEMISTRY DIVISION

Attention: DNA ANALYSIS LABORATORY
NATIONAL BUREAU OF INVESTIGATION
Taft Avenue, Manila 1000

DNA TYPING/PROFILING

It is an analytical procedure of identifying

the most likely owner of a piece of
evidence/biological sample.

To determine the genotype of the donor/source of the DNA material, the

NBI make use of the fully automated hi-tech equipment called ABI-PRISM
310 GENETIC ANALYZER and 3130 GENETIC ANALYZERS, using different
typing system.

ACCEPTABILITY IN COURT

The proposed Rule on DNA take effect on October 15,

2007 following its publication in a newspaper of
general circulation on October 2, 2007.
EN BANC A. M. NO. 06-11-5, SC
Rule on DNA Evidence Resolution

CASES RESOLVED CAUSE OF DNA ANALYSIS

1. Edgardo A. Tijing &n Bienvenida R. tijing v. Court of
Appeals and Angelita Diamante, G.R. 125901, March 08,
2001, (J. Quisumbing)
2. People v. Vallejo (G.R. No. 144656, May 09, 2002)
3. Carmelo Cabatania v. Court of Appeals & Carmelo Regodas, G. R. No.
124814, october 21, 2004
4. People of the Philippines v. Joel Yatar alias “Kawit” G.R. No.
150224, May 19, 2005, Per Curiam
5. People of the Philippines v. Rufino Umanito G.R. No 172607 April
16, 2009, (J. Quisumbing)
Directions: Submitted via FB Group trough the following format:

Student ID Number:
Student’s Name:
Permanent Address:
Year/Section:
Instructor:
School:

REVIEW QUESTIONS:
1. What is the role of serology in criminal investigation?

______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
2. What are the different methods employed in conducting blood
examination? Elaborate each method.

______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
3. What is DNA? What is its role in forensic investigation?

______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
4. Explain each the guidance for collecting and submitting
evidence.

______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
5. How does blood and DNA analysis makes crime investigation solve
complicated cases easily?
______________________________________________________________________________
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