Fk Unswagati Cerebro Spinal Fluid Analysis
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Transcript of Fk Unswagati Cerebro Spinal Fluid Analysis
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CEREBRO SPINAL FLUID
ANALYSISDEPARTMENT OF CLINICAL PATHOLOGY
MEDICAL SCHOOL- UNIVERSITAS PADJADJARAN/
RUMAH SAKIT DR. HASAN SADIKIN
BANDUNG
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Cerebrospinal fluid (CSF)
Produced 500 mL/ day
Produced by ultra filtration from blood plasma
and secreted by cells of the choroids plexus
water, circulates nutrients, and cushions andlubricates the central nervous system (CNS)
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Cerebrospinal fluid (CSF)
INCREASED AMOUNT :
acute and chronic congestion of the
meningens due to increasedtransudation; acute and chronic
infections
PRESSURE : 100-150 mm of water
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Sample collection of CSF:(PRE-ANALYTICAL STEPS)
CSF is collected by lumbar puncture
(between VL3-4 or VL4-5)
It is done only by definite indications:
1. DIAGNOSTIC or
2. THERAPEUTIC
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Sample collection of CSF:
(PRE-ANALYTICAL STEPS)
DIAGNOSTIC :
1.To study the CSF
2.To estimate intracranial pressure
3.To test the spinal block
4.To introduce air or lipoidal substance
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Sample collection of CSF:
(PRE-ANALYTICAL STEPS)..
THERAPEUTIC:1. To introduce penicillin, streptomycin, or
an anesthetic
2. To remove blood or irritative substance
CONTRAINDICATIONS :
1. Subtentorial tumors
2. Greatly increased intracranial pressure
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Sample collection of CSF:
(PRE-ANALYTICAL STEPS).
Freshly collected (within 30 minutes)
The amount of CSF to be collected :
8-10 ml
The first drop may contain blood from
the puncture and should not be used
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Sample collection of CSF:
(PRE-ANALYTICAL STEPS)
The specimen should be divided into
three sterile tubes, labeled 1,2, and 3:
1. chemistry and immunological exami-
nation
2. microbiological examination
3. cell count and differential count
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Sample collection of CSF:
(PRE-ANALYTICAL STEPS)..
When xanthochromic CSF is obtained,
add a trace of Lithium oxalate to
prevent clotting The cell count, bacteria and glucose
examinations must be done at once,
whilst the others can be delayed forseveral hours in the refrigerator
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ROUTINE ANALYSIS:
(ANALYTICAL STEPS)
Macroscopic examination:
Color, Normal: clear and colorless
Turbidity, Normal: clear
Reaction, Normal : alkaline
Specific Gravity, Normal : 1.003-1.008
Coagulation, Normal: does not coagulate
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Macroscopic examination:..
If the blood in the specimen is due to a
traumatic puncture, the CSF in the third
tube should be clearer than those in tube
1 or 2, after centrifugation, the superna-
tant should be clear
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Microscopic examination:
LEUKOCYTE CELL COUNT
DIFFERENTIAL COUNTINGInterpretation:
Normal only mononuclear (MN) cells
(lymphocytes or an occasional monocyte)
are found
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CHEMICAL EXAMINATION :
PROTEIN : Nonne test, Pandy`s test,and Total protein.
The main interest protein is globulin
Interfered by blood in CSF
CSF GLUCOSE :should be done at the
same time with serum glucoseexamination
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NONNE APELT TEST
Detection Globulin and albumin :
Is due to the blood interfering (Falsely high)
If the CSF is cloudy centrifuged the
clear supernatant can be used for the test
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NONNE-APELT TEST.
GLOBULIN
1cc CSF + 1cc ammonium sulfate (saturated)
Procedure : place the tip of pipette containing
amm.sulfate to the bottom of the test tube
which already contains with CSF. Let the
amm.sulfate solution layers underneath theCSF
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NONNE-APELT TEST..
Positive result : a clear-cut, grayish-white ring appears at the contact zone
between two fluids
Observe for 3 minutes. If a ring is formed,
then shake the tube to mix the fluids.Result should be report as follows :
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NONNE-APELT TEST.
+ = a ring appears within 3 min, only
visible against dark background,
leaving no trace on mixing
++ = a faint, opalescence after mixing
+++ = a definite cloud after mixing
++++ = a heavy cloud after mixing
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NONNE-APELT TEST
ALBUMIN
Shake the tube used in globulin test
Add 1 drop of 10% acetic acid, boil Report as follows:
Negative = a slight cloudiness
+ = a definite cloudiness, with a
fine precipitate
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NONNE-APELT TEST
++ = a flocculate precipitate in a slightly
cloudy fluid
+++ = a heavy flocculate precipitate in aclear fluid
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PANDYS TEST
1cc phenol (saturated aqueous solution) + 1
large drop of CSF
Normal CSF may show a faint trace, and
should be report as NEGATIVE
POSITIVE :
a bluish-white cloud forming immediately
around the drop of CSF :abnormal amount
of globulin
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CSF EXAMINATION :
GLUCOSE
DEPARTMENT OF CLINICAL PATHOLOGY
MEDICAL SCHOOL- UNIVERSITAS PADJADJARAN/
RUMAH SAKIT DR. HASAN SADIKIN
BANDUNG
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CSF GLUCOSE
Must be performed within 1.5 hrs after with-
drawl
Blood glucose: must be performed in thesame time
If possible, the blood and CSF are drawn
before breakfast
Normally : CSF glucose 60% blood glucose
level
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CSF GLUCOSE
Method : chemical & enzymatic
Chemicalmethods are no longer used,
because of lack of specificity, except ortho-
toluidine method
ENZYMATIC method :
Glucose oxidase (less specific than
hexokinase) Hexokinase (generally accepted reference
method)
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CSF GLUCOSE
GLUCOSE OXIDASE-PAP :
glucoseH2O
-D-glucose + O2 gluconolactone
oxidase O2peroxidase
gluconic acid + H2O2 H2O + OnOn + phenylamine-phenazone colorchanges
+ H2
O Measured by photometer in specific wavelength
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CSF GLUCOSE
HEXOKINASE :
hexokinase
Glucose + ATP glucose 6-phosphate + ADP
Mg++
G6PD
Glucose 6-phosphate + NADP 6 phosphoglucono-
lactone + NADPH + H+
More expensive, but better in specificity and precision
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CSF EXAMINATION :TOTAL PROTEIN
DEPARTMENT OF CLINICAL PATHOLOGY
MEDICAL SCHOOL- UNIVERSITAS PADJADJARAN/
RUMAH SAKIT DR. HASAN SADIKIN
BANDUNG
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PRINCIPLE OF THE TEST :
Protein in the presence of copper ions form
a violet blue color complex in alkaline solu-
tion (Biuret method)
The reagents used are same with reagents in
serum/plasma protein measurement
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REAGENTS :
R1 : Sodium hydroxide
Potassium sodium tartrate
R2 : Sodium hydroxidePotassium sodium tartrate
Potassium iodide
Copper sulphate
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The convention procedure, from serum/plasma
to CSF
The crucial things to do are :1. The standard solution must be diluted 100x
2. The amount of reagent and sample :
inversion from serum/plasma measurement(i.e. 20 L reagent + 1000 L CSF)
The photometer should be programmedfirst, then get the fixed FACTOR to calculate
the amount of protein
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INTERFERING FACTORS :
Ascorbic acid > 30 mg/dl
Bilirubin > 40 mg/dl
Hemoglobin> 500 mg/dl
Triglyceride > 1000 mg/dl
IV polydextrans : falsely too high levels
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Laboratory Activity
PATOLOGI KLINIK
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CEREBROSPINAL FLUID
Lumbar puncture between 3rd, 4th & 5th
lumbar vertebra
No special preparation
Freshly collected (30 minutes)
Chemistry
& serologymicrobiology
Hematology
& microbiology
1 2 3
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MACROSCOPIC
Color
Turbidity
ReactionSpecific Gravity
Coagulation
CELL COUNTING
RBC
WBC
ROUTINE ANALYSIS
CHEMISTRY
Glucose
Enzim (LDH, CK)
Protein
Qualitatitive: Nonne
& Pandy test
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NONNE TEST
Pipette 1 cc of
spinal fluid into a
small test tube
Gently add 1 cc of
ammonium sulfate
solution, to make 2
layers of solution
Positive reaction:
a clear cut, thin, grayish-white
ring appears at the zone of
contact of the two fluids within a
few second.
Normal : negative
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PANDY TEST
Add 1cc of phenol
solution to a tubeAdd 1 large drop of
spinal fluid
Normal:
Negative
Positive reaction:
bluish-white cloud
forming immediately
around the drop.