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Lab diagnosis of Meningitis
The document outlines the procedure for collecting and processing cerebrospinal fluid (CSF) samples for diagnosing meningitis caused by various pathogens, including bacteria, viruses, and fungi. It details the steps taken during a lumbar puncture, contraindications, sample handling, and various culture techniques required for diagnosis. It also discusses rapid diagnostic tests for identifying specific organisms through serological methods.
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Information on pyogenic, tubercular, aseptic infections, and examples of bacteria, fungi, and protozoa.
Overview of sample collection, transportation, initial processing, and various sample types including CSF and blood.
Step-by-step procedure for lumbar puncture, including patient consent, positioning, and CSF collection.
Guidelines for immediate processing of CSF samples, including centrifugation and analysis methods.
Diagnostic criteria for various types of meningitis based on cell counts, biochemical analysis, and CSF pressure.
Techniques used in direct microscopy, including dry smear and wet film preparations for pathogens.
Culturing methods for various pathogens from CSF samples, highlighting specific media and expected colony characteristics.Details on rapid diagnosis methods for meningitis, including serological techniques and specific tests for various pathogens.
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Lab diagnosis of Meningitis
- 2. Pyogenic Tubercular Aseptic Pyogenic Bacteria Eg. N.Meningitidis, S. pneumoniae, H. Influenzae Mycobacterium Tuberculosis Viral, Fungi and protozoa
- 3.
- 5. CSF – LumbarPuncture Blood – i.v. Other samples : For viruses : Nasopharyngeal & Throat swab, Feaces Cobweb appearance (Tubercular)
- 6.
- 7. Contraindications Local skininfections Suspected spinal cord or i/c mass lesion bleeding diathesis Spinal column deformities Non-cooperation of patient
- 8. Stylet Needle Black – 22G Yellow – 20 G Spinal Needle
- 9. STEP 1 PATIENT’S CONSENTIS TAKEN
- 10. STEP 2 PATIENT POSITIONING Laterallying position Sitting position
- 11. STEP 3 LANDMARKING PUNCTURESITE Line joining Post. Sup. iliac crests intersects Midline at Spinous process of L4
- 12. STEP 4 CLEANING SITEWITH ANTISEPTICS First Iodinated antiseptics Then, Isopropyl alcohol
- 13. STEP 5 LOCAL ANAESTHESIA 1%Lidocaine without Epinephrine
- 14. STEP 6 SPINAL NEEDLEINSERTION
- 16. STEP 7 CSF COLLECTIONIN 3-4 TEST TUBES TUBE 1 Glucose Protein TUBE 2 Gram stain Culture TUBE 3 Cell count TUBE 4 For special tests (2-5 mL each)
- 17. STEP 8 REMOVAL OFNEEDLE AND APPLYING BANDAID
- 19. Immediately hand-delivered Never refrigerated (H.Inf may die)
- 21. One PortionCentrifugated at 3000 rpm/5min Other portion left uncentrifuged SUPERNATANT DEPOSIT UNCENTRIFUGED CENTRIFUGED for Antigen detection, Biochemical Analysis for Direct Microscopy, Culture for Cell count
- 23. Cell Count Biochemical Analysis DirectMicroscopy Culture Antigen Detection
- 24. Diagnosis Total Cells (permm3) Predomina nt cell Proteins (mg/dL) Glucose (mg/dL CSF Pressure (mm Hg Normal CSF 0-5 Lymphocyte 15-45 40-80 70-180 Pyogenic Meningitis ↑↑ed (5-20,000) Polymorph (Neutrphl) ↑↑↑ed (>100) ↓↓ed ↑ed Tubercular Meningitis ↑ed (5-2,000) Lymphocyte ↑↑ed (>45) ↓ed ↑ed Viral Meningitis ↑ed (5-2,000) Lymphocyte ↑ed (>45) Normal ↑ed or Normal
- 25. Direct Microscopy Usingpart of sediment Two types : Dry Smear Wet film
- 26.
- 27. Direct Microscopy Wet Films AcanthamoebaCryptococcus
- 28. Culture CSF culture usingsediment For Pyogenic Bacteria : Blood Agar Macconkey Agar Chocolate Agar Brain-Heart Infusion Broth For M. tuberculosis : Lowenstein-Jensen (LJ) Middlebrook media For fungi : Sabouraud dextrose agar (SDA) Incubated at 37oC for 48-72 hours Incubated at 30oC for 8 weeks Incubated at 30oC for 3 weeks
- 29. Culture MacCconkey Agar (LF)Blood Agar (Str. Pneumoniae) Pink circular colonies Small, dome-shaped with greenish discolouration (α-hemolysis)
- 30. Culture Chocolate Agar (H. Influenzae) Brain-HeartInfusion Broth (Fastidious bacteria) Small opaque colonies
- 31. Culture Lowenstein-Jensen Medium (Egg based) Middle-brook Medium (Agarbased) Dry, Rough, Tough, buff coloured colonies Granular colonies
- 32. Culture SDA medium (Cryptococcus) Smooth,Mucoid, Cream- coloured colonies
- 33. Culture Blood Culture Usefulin meningitis due to : N. meningitidis H. influenzae Str. Pneumoniae Special Culture/isolation techniques If Viral/Protozoal/Fungal meningitis is suspected
- 34. Confirmatory Tests For confirmation Microscopy and biochemical reactions Serological techniques Other confirmatory tests depending on the organism
- 36. Rapid Diagnosis Supernatantfluid contains Antigen Detected Serologically Rapid Diagnostic Test for Meningitis Methods : Latex Agglutination, Counterimmunoelectrophoresis (CIEP)
- 37. Rapid Diagnosis Available for: N. meningitidis, Str. Pneumoniae, H. influenzae type b Group B streptococcus
- 38. Rapid Diagnosis Latex AgglutinationTest Slide AGGLUTINATION occurs Antigen Polystrene Latex Particles Antibody coated Supernatant
- 39. Rapid Diagnosis Counter-immuno-electrophoresis Antigen Antibodies Supernatant Slide Agarose layer Wellscut on surface - + Electric Current is applied PRECIPITATION LINE forms
- 40.