CHEMISTRY REAGENT KITS
           ------->CSF PROTEIN(Turbidometric) End-Point
                                             
                                                                          DOWNLOAD PDF

CLINICAL SIGNIFICANCE
Increased CSF Protein levels are the clinical Indication of various types of meningitis, polyneuritis and tumours. Increased Protein levels In urine are associated with Nephritis, Diabetic Malignancy or UTI.

PRINCIPLE
In the presence of sulphosalicylic acid and sodium sulphate, Proteins yield a uniform turbidity which absorbs maximum at 530 nM or GREEN filter and is directly proportional to the concentration of Proteins.

REAGENTS SUPPLIED
Cat. No. A192

Composition

 

1. CSFProtein Reagent

2x50 mL

  5-Sulphosalicylic Acid 120 mMol/L
  Sodium Sulphate 500mMol/L
2. Standard 15mL
  Albumin Fraction V 100 mGs/dL
 

Contains stabilisers and sodium azide as preservative.

Preparation of Working Reagent
All Reagents are ready to use.

STORAGE & STABILITY
Reagent No. 1,24 months at 25-35°C. Reagent No. 2,24 months at 2-8°C. DO NOT FREEZE STANDARD. Discard contaminated or turbid reagent.

SAMPLE
Non-traumatic CSF or Urine.
No prior patient preparation is required.
If urine to be assayed use fresh urine or add glacial acetic acid as a preservative to   a   final   concentration   of   0.1%   to   24 hour urine sample.
(All samples should be handled as potential infective agents as no laboratory methods make conclusive findings for its safety. Therefore, adequate protective laboratory measures should be taken while handling such materials).

SYSTEM PARAMETERS

Reaction

End-point

Temperature 25-35°C
Wavelength

530 nM (510-550 nM)

Secondary filter NIL
Standard Concentration 100mGs/dL
Absorbance Range 0-1 ° A
Cuvette Path Length 1 cM
Reagent Volume 500µL 1000µL
Sample Volume 20µL 50 µL
Reaction Time

0 Mins

Linearity 200 mGs/dL
Max. limit of Blank Reagent 0.050
Final Colour Stability 30 mins


METHOD FOR TOTAL CHOLESTEROL 1.
Pipette into 3 Test tubes Blank mL Standard mL Test mL
CSF Protein Reagent No 1 3.0 3.0 3.0
Distilled water 0.02 - -
Standard Reagent No 2 - 0.02 -
Sample (Urine or csf) - - 0.2

2.Mix well. Incubate at RT for 10 minutes.

3.Mix well and read at 530 nM (520-590 nM) or GREEN filter against Blank. Final reading must be taken within 30 minutes.

Micro method for discrete analysers

For Cuvette Volume 1.0mL 0.5mL
Working Reagent 1000µL 500µL
Sample or Standard 50µL 20µL

Mix well. Incubate at RT for 10 minutes. Mix well and read at 530 nM (510-550 nM) against Blank. Final reading must be taken within 30 minutes.

NOTE: Programme the analyser using system parameters. A specific programme data sheet may be provided for each analyser upon request.

RESULTS

                                           O.D.Test
Proteins in mGs/dL= -----------------------   x  100
                                         O.D.Standard

EXPECTED VALUES
CSF Proteins                             20-40 mGs/dL
Urine Proteins                20-140 mGs/24 hours

As with all diagnostic methods, the final
diagnosis should not be made on the result
of a single test as well as laboratory diagnosis must be confirmed with clinical manifestations.

LIMITATIONS
This assay is turbidometric and linear to 200 mGs/ dL Proteins. Globulin at higher concentrations may not obey Beer's Law. For levels higher than 200 mGs/dL. Proteins, repeat the test diluting the sample with 0.9% sodium chloride. Multiply the result with dilution factor, (i.e. 2 for a dilution of 1:1). Bichromatic reading will lead to false results.

Avoid Bichromatic reading.
It is appropriate to establish each laboratory's accuracy constant and interpret values accordingly. Similarly, laboratory findings should be established by clinical manifestations.

WARNING
This reagent system is for in vitro use only. This reagent system contains preservatives and components that have not established for safety if contacted on broken skin or eye or taken orally. In case of such incidents wash off with plenty of water, or consult a physician. concentrations and a graph be plotted.

QUALITY CONTROL
To ensure adequate quality control, each kit should be tested against a standard control sera. It should be realised that the use of quality control material checks both instrument and reagent function together. Factors which might affect the performance of this test include proper instrument function, temperature control, cleanliness of glasswares and accuracy of pippetting.

BIBLIOGRAPHY

  1. KING E.J. AND HASLEWOOD, GAD (1936), Lancet, 2,1153.
  2. KINGSBURG, F.P., CLARK C.P., WILLIAMS G. AND POST A.I. (1926) J. Lab. Clin. Med. 11981.