b iochemistry p ractical by: dr. beenish zaki. csf a nalysis
TRANSCRIPT
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BIOCHEMISTRY PRACTICALBy: Dr. Beenish Zaki
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CSF ANALYSIS
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CEREBROSPINAL FLUID
The surface of the central nervous system is covered by the meninges: Duramater Arachnoid Piamater.
The space between arachnoid and piamater is called sub-arachnoid space.
CSF is secreted by the choroid plexuses. In normal healthy adults rate of CSF formation is
100-250ml/24hrs.
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CSF FORMATION AND CIRCULATION
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SITE OF WITHDRAWL
•CSF is withdrawn by a procedure called lumbar puncture.•Spinal cord ends near the first lumbar vertebra.•CSF is obtained by passing Lumbar Puncture (L.P) needle between L4-L5.•Second preferred site is L3-L4 into the sub-arachnoid space.
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SITE OF LUMBAR PUNCTURE
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NORMAL COMPOSITION:
Colour : Clear colorless; no coagulum or deposit
PH : 7.3
Pressure:60-150 mm of Hg
Specific gravity: 1.006-1.002
Cells : 0-4 mononuclear cells/mm3
Protein content: 10-45mg/dl
Globulins: absent
Glucose: 45-100mg/dl
Chlorides: 120-130meq/l
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MEASURING OPENING PRESSURE
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ABNORMALITIES
In appearance
Blood stained a)Due to traumatic tap RBC under microscope
have normal shape b) In pathological conditions like sub-arachnoid
haemorrhage in this blood is homogeneously mixed with CSF under microscope RBC have a crenated appearence.
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ABNORMALITIES CONT’
In appearance Xanthochromia:-Yellow colorization of CSF
due to either: bilirubin Carotenoids
Turbidity :- is due to presence polymorphs/mm3
Coagulum:- In tuberculosis meningitis cob-web like coagulum is seen if the fluid is allowed to stand overnight from the web tubercle bacilli can be examined easily under microscope.
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ABNORMALITIES CONT’
In Glucose CSF glucose is increased in :- Diabetes Encephalitis Cerebral abscess CSF glucose is decreased in :- Meningitis due to bacteria. The glucose reduces from the fluid because
the organisms are glycolytic and thus use up the glucose.
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A CASE
A 19 year old student presents with fever, headache, nausea, vomiting. On
examination there is neck stiffness.
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INDICATION FOR CSF COLLECTION
CSF infection
Hemorrhage
Malignancy
Demyelinating Diseases
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CSF PROTEINS ESTIMATION
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METHOD AND PRINCIPLE
Method:Turbidity Method
Principle:Proteins are precipitated with Trichloro acetic acid which gives a turbidity.
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PROCEDUREProcedure Blank(ml) Test(ml)
Sample - 1.00
ReagentTCA
4.00 3.00
Mix well and take reading immediately using green filter
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OBSERVATION AND CALCULATION
O.D of Test= O.D of Standard=0.16 Concentration of sample =
O.D Test X concentration of standard (mg/dl)
O.D Standard
O.D Test x 100
0.16
O.D-Optical Density
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REFERENCE RANGE AND CLINICAL CONDITIONS
Adult: 10-45 mg/dl
CSF Protein Increased in: Meningitis Meningoma Acoustic Neuroma Multiple Sclerosis
CSF Protein Decreased in: Malnutrition
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CSF CHLORIDE ESTIMATION
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METHOD AND PRINCIPLE
Method: Titration Method
Principle:Chlorides present in CSF reacts with Silver nitrate to form silver chloride this reacts with the indicator to form a colored complex
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PROCEDURE
Pipette 1ml of CSF and 2ml of distilled water into a conical flask.
Add 3 drops of potassium chromate as the indicator.
Titrate against silver nitrate in the burette to a faint brick red color which is the end point.
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OBSERVATION AND CALCULATION
Initial Reading= Final Reading=
Concentration of sample:mEq. Chloride per liter of CSF= ml of silver nitrate solution required x 30
mEq/L= Milliequivalent/L
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REFERENCE RANGE AND CLINICAL CONDITIONS
Adult: 90-120 mEq/L
CSF Chloride Increased in: Hypertension Renal disease
CSF Chloride Decreased in: Meningitis
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THANK YOU!