ANALYSIS OF BODY FLUIDS

3A CEREBROSPINAL FLUID
Zairah F. Pascua | December 17, 2020

CEREBROSPINAL FLUID
 major fluid of the body which provides a
physiologic system
 considered as the 3rd major fluid in the body
 not an ultra filtrate of plasma
 abnormalities in composition of CSF diagnosed
diseases like: nncephalitis, meningitis, multiple
sclerosis, neurosyphilis, subarachnoid hemorrhage

CEREBROSPINAL FLUID
 First recognized by Cotugno (1794)

Functions:
1. To supply nutrients to the nervous tissue
 Acts as a medium/ transporter of nutrients
in the CNS
2. Remove metabolic wastes
 The one way flow from the CSF to the CSF is produced in the choroid plexuses of the two lumbar
blood takes potential harmful metabolites, ventricles and the third and fourth ventricles.
drugs and other substances away from
the pain Note: Adults- approx 20 mL of fluid is produced every hour.
3. Produce a mechanical barrier to cushion the brain
and spinal cord against trauma
 CSF protects and lubricates the brain Flows through the subarachnoid space located between
from damage/ trauma by buffering the the arachnoid and pia matter.
brain
 shock absorber
Reabsorbed back into the blood capillaries in the
The brain and spinal cord are lined by meninges which has arachnoid granulations/villae at a rate equal to its
3 Layers: production
a. Dura mater
 Latin for “hard mother” Note: To maintain a volume of:
 outermost layer that lines the skull and vertebral  Adults: 90 to 150 mL
canal  Neonates: 10 to 60 mL in neonates
b. Arachnoid mater
 “spiderweb-like” BLOOD-BRAIN BARRIER
 middle layer, filamentous (spider-like) inner  selective barrier
membrane.  used to represent the control and filtration of blood
c. Pia mater components to the CSF and then to the brain
 Latin for “gentle mother”  Composed of capillary endothelium held together
 innermost layer, thin membrane lining the by tight junction and choroid plexuses
surfaces of the brain and spinal cord  Allows essential metabolites (O2, glucose) to pass
Note: from the blood to the CNS
Subarachnoid space- between arachnoid and pia matter  Blocks most molecules >500 daltons
- contains CSF  Damage to the blood-brain barrier allows entrance
of increase amounts of proteins and leukocytes;
 also causes disease meningitis and multiple
sclerosis

SPECIMEN COLLECTION AND HANDLING

METHODS OF COLLECTION:
Lumbar Puncture
 routine method for CSF collection
 between 3rd, 4th or 5th lumbar vertebrae
 patient is in fetal position or lateral
decubitus position
 Although this procedure is not
complicated, it does require certain
precautions:
a. measurement of intracranial pressure

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b. careful technique to prevent infection  Tube 3- for cell count/ Hematology

or neural tissue damage
 The volume of CSF that can be removed
is based on the volume available in the
patient (adult vs. neonate) and the
opening pressure of the CSF, measured
when the needle first enters the
subarachnoid space.
Take Note:
 Adults- between 3rd or 4th lumbal vertebrae
-least likely to contain cells introduced by the
spinal tap procedure
-refrigerated at 2-8℃
 Tube 4- drawn for the microbiology laboratory for
better exclusion of skin contamination or for
additional serologic tests
Note:
 Low volume CSF specimen collected from 1 tube-
order of testing should be:
a. Microbiology
b. Hematology
c. Chemistry/ Serology
 Excess fluids should not be discarded, should be
frozen until there is no further use for it.

NORMAL VALUES FOR CSF

 Color: colorless
 Viscosity: same with water
 Clarity: Crystal clear
 Specific gravity: 1.006-1.008
 pH: 7.30-7.45
 Pressure: 50-200 mmH2 O

Examination of the CSF occurs first at the bed side. It is

also included in the laboratory report.

Terminology used to describe CSF appearance:

 Children- between 4th or 5th lumbar vertebrae APPEARANCE CAUSE MAJOR
SIGNIFICANCE
Slightly Hazy WBCs Meningitis
Other Methods: 200-500/microliter
1. Ventricular Puncture Cloudy/ Turbid/ WBCs (above Meningitis
 used in infants with open fontanels Milky 500/microliter) Disorders affecting
2. Cisternal Puncture microorganisms BBB
 collected in the sub-occipital region increased proteins Production of IgG
3. Lateral Cervical Puncture or lipid within the CNS
concentration
Specimens are collected in 3 sterile tubes: Clotted Increased proteins Froin’s disease
and clotting
ORDER OF PRESERVATION factors
COLLECTION Bloody RBCs Non-pathologic:
Tube 1 Chemistry and Frozen traumatic tip
Serology Pathologic:
Tube 2 Microbiology Room hemorrhage
temperature Pellicle Increased proteins Tubercular meningitis
Tube 3 Hematology/ Cell Refrigerated formation and clotting
count factors
Note: Xanthochormic Hemoglobin Old
Note: Ideally, CSF test is performed on STAT basis. If hemorrhage/lysed cell
this is not possible, specimens are maintained by from traumatic tap
preservation. Bilirubin RBC
 Tube 1 - for chemical and serologic tests degradation/elevated
-these tests are least affected by blood or serum bilirubin levels
bacteria introduced as a result of the tap Carotene Increased serum
procedure. levels
-frozen at -15 to -30℃ Protein Disorders affecting
BBB
 Tube 2- usually designated for microbiology Melanin Melanin
laboratory melanosarcoma
-remains at room temperature at 19-26 Oily Radiographic
degrees contrast media

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Note: CHEMICAL EXAMINATION

 Fluid for centrifugation must be in capped tubes. Note: Reference values for CSF Chemicals are not the
 All specimens should be treated with extreme care same as the plasma values because the filtration process is
because they can be highly contagious. selective; the chemical composition is controlled by the
Blood-Brain Barrier.
Xanthochromia
 term used to describe CSF supernatant that is pink, I. CEREBROSPINAL PROTEIN
orange, or yellow
 most common cause: presence of RBC degradation  most frequently performed chemical test on CSF
products
 Depending on the amount of blood and the length of Normal values: 15-45 mg/dL
time it has been present, the color will vary from:  not reported in g/dL like in plasma
1. Pink- very slight amount of oxyhemoglobin) to *values are higher in infants and older persons
2. Orange- heavy hemolysis
3. Yellow- conversion of oxyhemoglobin to
unconjugated bilirubin PROTEINS NORMALLY FOUND IN CSF:
Note: 1. Albumin- major CSF protein
Other causes of xanthochromia: 2. Prealbumin- second most prevalent fraction in CSF
1. Elevated serum bilirubin - alpha globulins include primarily
o Xanthochromia caused by bilirubin is haptoglobin and ceruloplasmin
commonly seen in premature infants. 3. Haptoglobin and Ceruloplasmin
2. Presence of the pigment carotene 4. Transferrin- major beta globulin present
3. Markedly increased protein concentrations, and  Tau protein- separates carbohydrate-deficient
melanoma pigment transferrin fraction seen only in CSF, not in
serum.
4. IgG with small amounts of IgA
TRAUMATIC COLLECTION DIFFERENTIATION
- CSF gamma globulin is primarily
immunoglobulin G (IgG), with only a small
amount of immunoglobulin A (IgA)
DIFFERENCES INTRACRANIAL TRAUMATIC
HEMORRHAGE TAP PROTEINS NOT NORMALLY FOUND IN CSF:
Distribution of blood Even in all 3 tubes Uneven 1. IgM
Clot formation Absent Present 2. Fibrinogen
Xanthochromic Common Not common 3. Beta lipoprotein
supernatant
Erythrophagocytosis Present Absent Clinical Causes of Abnormal CSF Protein Values*
/ Hemosiderin
granules
D-dimer Test Positive Negative
Note:
 Traumatic tap: tube 1 has the heaviest concentration
of blood and gradually diminishing amounts in Tubes 2
and 3.
 Clot formation: Hemorrhage doesn’t clot because it
doesn’t contain enough fibrinogen to clot.
Traumatic tap may form clots owing to the introduction
of plasma fibrinogen into the specimen

Increase proteins and clotting factors can also cause

clot formation but do not usually produce blood fluid in
conditions:
a. Meningitis
b. Froin syndrome
c. Blocked CSF circulation
 RBCs must usually remain in the CSF for approx. 2
hours before noticeable hemolysis begins; therefore, a
xanthochromic supernatant would be the result of Note:
blood that has been present longer than that  Meningitis and hemorrhage conditions that damage
introduced by the traumatic tap the blood–brain barrier- most common causes of
To examine a bloody fluid for the presence of elevated CSF protein.
xanthochromia, the fluid should be centrifuged in a
microhematocrit tube and the supernatant examined
against a white background. METHODS FOR CSF PROTEIN
I. QUALITATIVE TEST

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TEST REAGENT POSITIVE RESULT  The index increases relative to the amount of
Nonne-Apelt Ammonium Cloudy precipitate damage to the barrier.
sulfate
Rose Jones Ammonium White ring CSF/serum albumin index = CSF albumin (mg/dL)
sulfate Serum albumin (g/dL)
Pandy’s Phenol Blueish white cloud
Nogochi 10% butyric Precipitate CSF IgG Index
acid  measures IgG synthesis within the CNS
Colloidal Gold Test Colloidal gold 1+ (blue color)  Index values > 0.70- indicate IgG production
solution 2+ (purple color) within the CNS
3+ (deep blue color)
4+ (pale blue color) IgG index = CSF IgG (mg/dL)/serum IgG (g/dL)
5+ (colorless) CSF albumin (mg/dL)/serum albumin (g/dL)
II. QUANTITATIVE TESTS: 5. ELECTROPHORESIS
2 most routinely used techniques for measuring total  To detect oligoclonal bands
CSF protein use the principles of  The primary purpose for performing CSF
 turbidity production or dyebinding ability. protein electrophoresis is to detect
oligoclonal bands, which represent
1. TURBIDIMETRIC inflammation within the CNS
 precipitation of protein using:  Agarose gel electrophoresis followed by
 Trichloroacetic acid (TCA) Coomassie brilliant blue staining- most
frequently performed in the clinical
- reagent of choice
laboratory.
- precipitates both albumins  Better resolution can be obtained using
and globulins CSF immunofixation electrophoresis
 Sulfosalicylic Acid (SSA) (IFE) and isoelectric focusing (IEF)
- precipitates albumin only; followed by silver staining. These
unless combined with sodium techniques are the method of choice when
sulfate determining whether a fluid is actually CSF
Note:
2. DYE-BINDING TECHNIQUE Electrophoresis- method of choice when it is
 Principle: Protein error of indicators necessary to determine if the fluid is CSF- “Tau
 Advantages: identification”
1. requires smaller sample size
2. less interference from external Multiple Sclerosis
surface  presence of two or more oligoclonal bands
 Stains used: in the CSF that are not present in the serum
 Coomassie Brilliant Blue G250- binds part when accompanied by an increased
to variety of proteins IgG index
- color change: from red to blue  Oligoclonal banding remains positive during
- uses beer’s law remission of multiple sclerosis, but
 Ponceau S disappears in other disorders.
3. NEPHELOMETRY
 Uses benzalkonium chloride Other Disorders with Oligoclonal bands but not
4. PROTEIN FRACTIONS in serum:
 To accurately determine whether IgG is a. Encephalitis
increased because it is being produced b. Neurosyphilis
within the CNS or is elevated as the result c. Guillain-Barré syndrome
of a defect in the blood–brain barrier, d. Neoplastic disorders
comparisons between serum and CSF
levels of albumin and IgG must be made. 6. MYELIN BASIC PROTEIN (MBP)
 diagnosis of neurologic disorders  presence of myelin basic protein (MBP) in
associated with abnormal CSF protein the CSF indicates recent destruction of the
often requires measurement of the myelin sheath that protects the axons of the
individual protein fractions neurons (demyelination)
 used to monitor course of multiple sclerosis
CSF/ Serum Albumin Index  provide a valuable measure of the
 To evaluate the integrity of the blood-brain effectiveness of current and future
barrier treatments
 calculated after determining the concentration  detected by radio immunodiffusion
of CSF albumin in milligrams per deciliter and
the serum concentration in grams per II. CSF GLUCOSE
deciliter. Normal value: 60-60% plasma glucose
 Index value < 9- intact blood– brain barrier

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 Glucose enters the CSF by selective transport 1. Liver disorder that results in increased blood
across the blood– brain barrier ammonia and CSF ammonia
 blood glucose should be drawn about 2 hours before 2. Comma of unknown origin
the spinal tap to allow time for equilibration between 3. Reye’s syndrome
the blood and fluid. o Approximately 75% of children with Reye
 Specimens should be tested immediately because syndrome have elevated CSF glutamine
glycolysis occurs rapidly in CSF levels
Notes: CSF: ASAP (within 30 minutes) Disturbance in consciousness occurs when levels are
>35 mg/dL
CLINICAL SIGNIFICANCE: TABLE 10-6 MAJOR LABORATORY RESULTS FOR
INCREASED: DIFFERENTIAL DIAGNOSIS OF MENINGITIS
1. Results of plasma glucose elevations
2. DM, encephalitis and conditions associated with 4 TYPES OF MENINGITIS:
intracranial pressure
1. Bacterial Meningitis
neutrophil is present
DECREASED:
>35 mg/dL lactate level
1. Aids in determining the causative agent of
2. Viral Meningitis
Meningitis
only lymphocyte is present
o Bacterial meningitis- markedly
3. Tubercular Meningitis
decreased CSF glucose level both lymphocytes and monocytes are present
accompanied by an increased WBC pellicle formation
count and a large percentage of decreased glucose level
neutrophils 4. Fungal Meningitis
2. Alterations in the mechanisms of glucose transport both lymphocytes and monocytes are present
across BBB Positive India ink and immunologic test for C.
3. Increased glucose use by brain cells neoformans
Note: normal to decrease glucose level
Tubercular meningitis - if the WBCs are lymphocytes
instead of neutrophils
Result:
Viral meningitis - if a normal CSF glucose value is WBC count- all are elevated
found with an increased number of lymphocytes

III. CSF LACTATE

Normal value: 10-24 mg/dL
 can be a valuable aid in diagnosing and managing
meningitis cases
 CSF lactate levels remain elevated during initial
treatment but fall rapidly when treatment is
successful, thus:
a sensitive method for evaluating the effectiveness
of antibiotic therapy
 used to monitor severe head injuries
 RBCs contain high concentrations of lactate
 falsely elevated results may be obtained on
xanthochromic or hemolyzed fluid

IV. CSF GLUTAMINE

Normal value: 8-18 mg/dL
 produced from ammonia and α -ketoglutarate by the
brain cells. This process serves to remove the toxic
metabolic waste product ammonia from the CNS
 chemical test frequently performed in CSF but not in
blood
 used an indirect measure of CSF ammonia
 preferred over the direct measurement of CSF
ammonia because the glutamine concentration
remains more stable than the volatile ammonia
concentration in the collected specimen
*As CSF ammonia increase- alpha-ketoglutarate
decreases= coma

ELEVATED:

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LD ISOENZYMES  Pasteur pipet is used to load the

 help diagnose meningitis by confirming the hemocytometer
presence of PMN and lymphocytes  Clear specimens may be counted
Increased LD Isoenzymes Condition undiluted, provided no overlapping of cells
LD1 and LD2 Brain tissue destruction
LD2 and LD3 Viral meningitis
LD4 and LD5 Bacterial meningitis

MICROSCOPIC EXAMINATION
I. CELL COUNT
 RBC, WBC, Total cell count
 Specimens that contain up to 200 WBCs or 400
RBCs/ µL may appear clear so it is necessary to
examine all specimens microscopically.
 Any cell count should be done immediately (within
30 minutes)
 WBC and RBCs begin to lyse within 1 hr.
is seen during the microscopic examination
o particularly granulocytes  Diluent used: Normal Saline Solution
 40% of leukocytes disintegrate after 2 hrs Neubauer counting chamber
Note: Specimens that cannot be analyzed immediately  3mm by 3mm
should be refrigerated  9 medium square
o 1 mmx1mm each
1. WBC COUNT
WBC count
 cell count routinely performed on CSF  corner squares- bec. WBC conc is lower than
specimens
RBC; a larger square is required to perform the
 lysis of RBCs must be obtained before cell count
performing the WBC count on either
o 16 smaller squares
diluted or undiluted specimens
width: 0.25mm
o use 3% glacial acetic acid to lyse
area: 0.0625 mm2
RBCs RBC, Platelet count
o Methylene blue is added to stain
 central square (5)- for platelets and RBCs
the WBCs providing a better  25 squares
differentiation between width: 0.2mm
neutrophils and mononuclear area: 0.04 mm2
cells  16 smaller squares
 Normal CSF WBC count: width: 0.05 mm
 Adult: 0-5 WBC/microliter area: 0.0025mm2
 Children: values are higher than adult
 Neonates: 0-30 mononuclear COUNTING CHAMBERS USED:
cells/microliter
 Dilution used depends on clarity of  cells counts in CFS are performed in a manual
specimen: counting chamber:
CLARITY DILUTION
1. Improved Neubauer Counting Chamber
Slightly hazy 1:10
 routinely used for performing CSF cell counts
Hazy 1:20
Slightly cloudy 1:100 # of cells x Dilution = cells/µL
Slightly bloody or 1:200 # of cells counted x Vol. of 1 square
Cloudy
Bloody or Turbid 1:10,000 2. Fuchs-Rosenthal Counting Chamber
 undiluted specimen; phase microscopy
2. RBC COUNT  16 large squares; depth=0.2 mm
 no RBC should be present in normal CSF  Cells per microliter= # of cells
 done only when there is traumatic tap and 3
correction for leukocytes or proteins is Note:
needed Electronic cell counters - not been used for performing
 RBC count= Total cell count- WBC count CSF cell count bec of high background counts

3. TOTAL CELL COUNT The standard neubauer calculation formula used blood cell
 Cells are counted in the four corner count is also applied to CSF cell counts.
squares and the center square on both This formula can be used for both undiluted and diluted
sides of the hemocytometer. specimens.

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QUALITY CONTROL OF CSF AND OTHER BODY FLUID  lymphocyte: monocyte (70:30)
CELL COUNTS 2. Monocytes
 All diluents should be checked biweekly for  predominant in children
contamination by examining them in a counting  ratio 30:70
chamber under 400× magnification. 3. Neutrophils
 Contaminated diluents should be discarded and  rarely seen
new solutions prepared. Pleocytosis- increased numbers of normal cells and
 The speed of the cytocentrifuge should be checked considered as abnormal finding
monthly with a tachometer  When pleocytosis involving neutrophils,
 The timing should be checked with a stopwatch. lymphocytes, or monocytes is present, the CSF
 If nondisposable counting chambers are used, they differential count is most frequently associated with
must be soaked in a bactericidal solution for at its role in providing diagnostic information about
least 15 minutes and then thoroughly rinsed with the type of microorganism that is causing an
water and cleaned with isopropyl alcohol after each infection of the meninges (meningitis)
use.
ABNORMAL:
II. DIFFERENTIAL COUNT 1. Immature WBCs
 specimens should be concentrated and 2. Eosinophis
performed on a stained smear 3. Plasma cells
 Identifying the type or types of cells present in 4. Macrophages
the CSF is a valuable diagnostic aid. 5. Increased tissue cells and malignant cells
 100 cells should be counted, classified and
reported in percentage
 If <100 cells= report only the numbers of 1. NEUTROPHILS
the cell types seen  contain cytoplasmic vacuoles following
cytocentrifugation
 present in bacterial meningitis; and in the
METHODS OF SPECIMEN CONCENTRATION early stages (1 to 2 days) of viral, fungal,
1. Sedimentation tubercular, and parasitic meningitis
2. Filtration  Increased
 Sedimentation and Filtration- not routinely used a. central nervous system (CNS)
but produces less stellular distortion hemorrhage
b. repeated lumbar punctures
3. Cytocentrifugation c. injection of medications or
 As little as 0.1 mL of CSF combined with radiographic dye
one drop of 30% albumin produces an  pyknotic nuclei- indicate degenerating
adequate cell yield cells; may resemble nucleated red blood
 Adding albumin increases the cell yield cells (NRBCs) but usually have multiple
and decreases the cellular distortion nuclei
frequently seen on cytocentrifuged  NRBCs are seen as a result of
specimens bone marrow contamination
ex: during the spinal tap
a. cytoplasmic vacuoles
b. nuclear clefting 2. LYMPHOCYTES
c. prominent nucleoli  Reactive lymphocytes containing
d. indistinct nuclear increased dark blue cytoplasm and
e. cytoplasmic borders clumped chromatin are frequently present
f. cellular clumping that resembles during viral infections in conjunction with
malignancy normal cells
4. Centrifugation  mixture of lymphocytes and monocytes is
 specimen is centrifuged for 5-10 minutes common in cases of viral, tubercular, and
 supernatant fluid is removed and stored fungal meningitis
for additional tests  Increased lymphocytes are seen in cases
 slides made from the suspended of both asymptomatic HIV infection and
sediment are allowed to air dry and are AIDS
stained with Wright’s stain.  A moderately elevated WBC count (<50
WBCs/µ L) with increased normal and
reactive lymphocytes and plasma cells
may indicate multiple sclerosis or other
CSF CELLULAR CONSTITUENTS degenerative neurologic disorders.
NORMAL:
3. EOSINOPHILS
1. Lymphocytes
 predominant in adults  Increased eosinophils are seen in the CSF in
association with:
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a. parasitic infections c. renal

b. fungal infections (primarily Coccidioides d. gastrointestinal malignancies
immitis)  primarily from lung, breast, renal, and
c. introduction of foreign material, including gastrointestinal malignancies
medications and shunts, into the CNS  Cells from primary CNS tumors include
astrocytomas, retinoblastomas, and
4. MACROPHAGES medulloblastomas
 Purpose: remove cellular debris and foreign  usually appear in clusters and must be
objects such as RBCs distinguished from normal clusters of ependymal,
 appear within 2 to 4 hours after RBCs enter the choroid plexus, lymphoma, and leukemia cells.
CSF and are frequently seen following repeated
taps TABLE 10-3: PREDOMINANT CELLS SEEN IN CSF
 finding of increased macrophages indicates a
previous hemorrhage

NONPATHOLOGICALLY SIGNIFICANT CELLS MICROBIOLOGY EXAMINATION

 most frequently seen after diagnostic procedures
such as pneumoencephalography and in fluid
obtained from ventricular taps or during
neurosurgery
 appear in clusters and can be distinguished from
malignant cells by their uniform appearance
1. CHOROIDAL CELLS
 from the epithelial lining of the choroid
plexus
 seen singularly and in clumps
 Nucleoli are usually absent and nuclei
have a uniform appearance
2. EPENDYMAL CELLS
 from the lining of the ventricles and
neural canal
 have less defined cell membranes and
are frequently seen in clusters
 Nucleoli are often present
3. SPINDLE-SHAPED CELLS
 represent lining cells from the arachnoid
 usually seen in clusters and may be seen
with systemic malignancies
MALIGNANT CELLS OF HEMATOLOGIC ORIGIN
 Lymphoblasts, myeloblasts, and monoblasts in the
CSF are frequently seen as a serious complication
of acute leukemias
 Nucleoli are often more prominent than in blood
smears.
1. LYMPHOMA CELLS
 also seen in the CSF and indicate
dissemination from the lymphoid tissue
 resemble large and small lymphocytes
and usually appear in clusters of large,
small, or mixed cells based on the
classification of the lymphoma
 nuclei may appear cleaved, and
prominent nucleoli are present
Role of microbiology laboratory in analyzing CSF: identify
the causative agent in meningitis.
Several methods available to provide information for a
preliminary diagnosis:
1. GRAM STAIN
 recommended method for detection of
microorganism
 Use of the cytocentrifuge provides a highly
concentrated specimen for Gram stains
 routinely performed on CSF from all
suspected cases of meningitis
 All smears and cultures should be performed
on concentrated specimens
 CSF should be centrifuged at 1500 gravity for
15 minutes
 Presence of 105 per mL of organism is
needed for the initiation by gram stain
 at least a 10% chance exists that Gram stains
and cultures will be negative
Note:
 CSF Gram stain is one of the most difficult slides to
interpret because the number of organisms present
is usually small, and they can easily be overlooked,
resulting in a falsenegative report.
 false-positive reports can occur if precipitated stain
or debris is mistaken for microorganisms
ORGANISMS MOST FREQUENTLY ENCOUNTERED:
GRAM (+)
 S. pneumoniae
 common in adults
 S. agalactiae and L. monocytogenes (newborns)
GRAM (-)
 H. influenza
 E.coli common in adults
 N. meningitidis

2. BLOOD CULTURE
MALIGNANT CELLS OF NONHEMATOLOGIC ORIGIN  done because the causative agent will
 Metastatic carcinoma cells of nonhematologic be in both blood and CSF
origin are primarily from :
a. lung 3. ACID FAST/ FLUORESCENT ANTIBODY
b. breast STAINS

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 not routinely used  for the diagnosis of tertiary syphilis through the
 for Tubercular meningitis demonstration of antibodies associated with
syphilis
4. INDIAN INK 1. Venereal Disease Research Laboratories (VDRL)
 detects the presence of Cryptococcus  recommended for CSF by CDC to
neoformans diagnose syphilis due to its ability to detect
 C. neoformans- produce starburst active cases of syphilis
pattern in gram stain
 not as sensitive as FTA-ABS absorption test for
syphilis
5. LATEX AGGLUTINATION TEST AND ELISA
 provide a rapid means for detecting and
identifying microorganisms in CSF 2. Rapid plasma regain (RPR)
 detect:  test is not recommended because it is less
1. Streptococcus group B sensitive than the VDRL
2. H. influenzae Type B
3. S. pneumoniae
4. N. meningitidis A, B, C, Y, W135 3. Fluorescent Treponemal Antibody-absorption (FTA-
5. Mycobacterium tuberculosis ABS)
6. Coccidioides immitis  used with care to avoid contamination with
7. E. coli K1 antigens blood because it remains positive in the serum
of treated cases of syphilis
6. BACTERIAL ANTIGEN TEST (BAT)
 does not appear to be as sensitive to N.
meningitidis as it is to the other organisms
 should be used in combination with
results from the hematology and clinical
chemistry laboratories for diagnosing
meningitis

7. REVERSE LATEX AGGLUTINATION

 detects C. neoformans antigen in
serum and CSF
 more sensitive than indian ink
 False (+): Interference of rheumatoid
factor
 False (+) is inhibited by incubation
with:
a. dithiothreitol or pronase
b. boiling with ethylenediaminetetra-
acetic acid (EDTA)

8. LATERAL FLOW ASSAY (LAF)

 provide a rapid method for detecting
C. neoformans

9. LIMULUS LYSATE TEST

 diagnosis of meningitis caused by
gram (-) organisms by detecting
endotoxins found in their cell walls
 uses blood cells of the horseshoe
crab termed as “amebocytes” which
contains copper complex responsible
for the blue color.
 coagulates the amebocyte lysate
within 1 hour at 37℃
 sensitive to minute amounts of
endotoxin and will detect all gram
negative bacteria

SEROLOGIC EXAMINATION
 to detect the presence of neurosyphilis

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