LABORATORY DIAGNOSIS

MENINGITIS
SPECIMENS: CSF,BLOOD
CSF : By Lumbar puncture (or Spinal tap in L3-L4 level)

-Appearance of the CSF should be noted and recorded as: clear, hazy, turbid purulent, yellow to
xanthochromic (due to haemolysis or icterus), blood- tinged, with fibrin web or pellicle.
-Quantity: A minimum of 5 to 10 ml is reccomended (less can lead to false – results)
( Larger volumes 10-15 ml for Mycobacterium tuberculosis)

CSF is commonly collected into three/four tubes:

-1 Tube: It is used for chemistry studies ,glucose ,protein count and immunology studies
-2 Tube: It is used for culture
-3 Tube: It is used for cell count (Examined for WBC,RBC,protein content and glucose level)
-4 Tube: It is used for cytology

After collectionCSF shoulf be analyze immediately to the laboratory.

-CSF should be mantained at room T (S.pneumonia not be detectable after 1 h or longer)
-Refrigeration not recommended (exeption if involves viral studies)
In people with bacterial meningitis CSF often shows:
-turbid purulent appearance of CSF
-"opening pressure" of the CSF is elevated
-a low sugar (glucose) level
-increased WBC count(predominantly neutrophils)
- increased protein.
The type of WBC predominantly present and the appearance of the CSF predicts whether
meningitis is due to bacterial or viral infection.

 Normal CSF, mildly xanthochromic CSF, moderately xanthochromic CSF, red-tinged turbid CSF caused by
hemorrhage, and cloudy red-tinged fluid from a horse with bacterial meningitis.
Contraindications of lumbar puncture
if there is a mass in the brain (tumor or abscess) or the intracranial pressure (ICP) is elevated, as it
may lead to brain herniation. If someone is at risk for either a mass or raised ICP (recent head
injury, localizing neurological signs), a CT or MRI scan is recommended prior to the lumbar
puncture. In case of contraindication + we use other means for diagnosis like blood culture.

DIRECT MICROSCOPICAL EXAMINATION

If CSF is purulent (very cloudy) it can be examined without centrifugation.
In other cases, the CSF should be centrifuge in the laboratory and smears are prepared from the
sediment.

-Gram stain
Allows early identification and is low sensitive
In case of acute bacterial meningitis these stains reveal numerous WBCs and bacteria with specific
morphology ( N.Meningitidis, S.Pneumonia, H.Influenzae)
- Ziehl-Neelson stain is indicated to demonstrate AFB(Acid-Fast Bacillus) along with
LYMPHOCYTE in case tuberculous meningitis is suspected.
-India ink wet mount is recommended in cases suspected for fungal meningitis such as
C.neoformans for demostration of the polysaccharide capsule of organisms.
(if affects patient with AIDS culture is essential).
DIRECT ANTIGEN DETECTION TESTS
Detection of antigen in CSF may accomplish by techniques of latex agglutination or
coagglutination tests. These tests use the principle of an antibody-coated particle that will bind to
specific antigen, resulting in macroscopically visible agglutination. The capsular polysaccharide
antigens of most common etiological agents of meningitis are used.

MOLECULAR METHODS
-Highly specific and sensitive for CNS infection caused by herpes simplex virus and enteroviruses
-PCR reccomended to diagnosis of viral meningitis

MISCELLANEOUS TESTS
Other tests: limulus lysate test,CSF lactate determinations,C-reactive protein,mass spectrometry and
gas liquid chromatography

CULTURES
-Higher sentitive
for bacteria
-Centrifuged CSF sediment should be inoculated onto chocolate agar, Blood agar plate, enrichment
broth (thioglycolate) and incubated at 37C in 5/10% CO2 for at least 72 hours.
-S.pneumonia recognition occurs in blood agar plate
-Anaerobic bacteria from cerebral brain abscesses an anaerobic blood agar plate may be inoculated
-For CSF fungal and M. tuberculosis culture should be inoculated onto special media,Sabouraud
dextrose agar at 30 °C for 4 weeks

BRAIN ABSCESS
SPECIMENS: abscess material
-Shoul be submitted under anaerobic conditions
-Inoculated into sheep blood agar and chocolate agar plate
-Incubation in 5 /10% CO2 for 72 h at 35 C°