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CSF Protein

This document provides instructions for quantitatively determining the total protein levels in urine and cerebrospinal fluid (CSF) samples using a pyrogallol red colorimetric method. Proteins react with pyrogallol red and molybdate in an acid solution to form a colored complex, and the intensity of the color is proportional to the protein concentration. Elevated protein levels in urine can indicate renal disease, while elevated CSF protein may be caused by infections or increased intracranial pressure. The test has a measurement range of 9.44-4000 mg/L and is precise with intra- and inter-assay CVs below 5% and 16% respectively.

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78 views1 page

CSF Protein

This document provides instructions for quantitatively determining the total protein levels in urine and cerebrospinal fluid (CSF) samples using a pyrogallol red colorimetric method. Proteins react with pyrogallol red and molybdate in an acid solution to form a colored complex, and the intensity of the color is proportional to the protein concentration. Elevated protein levels in urine can indicate renal disease, while elevated CSF protein may be caused by infections or increased intracranial pressure. The test has a measurement range of 9.44-4000 mg/L and is precise with intra- and inter-assay CVs below 5% and 16% respectively.

Uploaded by

Phong Nguyễn Windy
Copyright
© © All Rights Reserved
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PROTEIN u&csf

Protein in urine and CSF


Pyrogallol red. Colorimetric

Quantitative determination of total urinary and CSF SPIN640 APPLICATION


protein
IVD
Store at 2-8ºC
PRINCIPLE OF THE METHOD
Proteins react in acid solution with pirogallol red and molybdate to
form a colored complex.
The intensity of the color formed is proportional to the protein
concentration in the sample1,2.

CLINICAL SIGNIFICANCE
In healthy persons, the urine contains no protein or only a trace
amount of protein; normally the glomeruli prevent passage of
protein from the blood to the glomerular filtrate. Glomerular injury
causes increased permeability to plasma proteins, resulting in
proteinuria, which refers to the presence of protein in the urine. SPIN640Plus APPLICATION
A persistent finding of proteinuria is the single most important
indication of renal disease.
Elevated concentration of protein in cerebro-spinal fluid (CSF) can
be cause by infections and intracranial pressure1,5,6.
Clinical diagnosis should not be made on a single test result; it
should integrate clinical and other laboratory data.

REAGENTS
Pyrogallol red 50 μmol/L
R
Sodium molybdate 0,04 mmol/L

PREPARATION The Calibration is stable until 36 days. After this period the Calibration
The reagent is ready to use. must be performed again in order to obtain good results.

STORAGE AND STABILITY PERFORMANCE CHARACTERISTICS


All the components of the kit are stable until the expiration date on Measuring range: From detection limit 9,44 mg/L to linearity limit of
the label when stored tightly closed at 2-8ºC protected from light 4000 mg/L.
and contaminations prevented during their use. If the concentration is greater than linearity limit, dilute the sample 1/2
Do not use reagents over the expiration date. with NaCl 9 g/L and multiply the result by 2.
Precision:
Signs of reagent deterioration: Intra-assay (n= 20) Inter-assay (n= 20)
 Presence of particles and turbidity. Mean (mg/L) 220 536 1014 216 499 1018
 Blank absorbance (A) at 600 nm  0,30. SD 3,7 4,0 5,2 18,3 26,1 166,1
CV (%) 2,28 0,75 0,51 7,35 5,22 16,43
ADDITIONAL EQUIPMENT Sensitivity: 1mg/L = 0,00026 (A).
 SPIN640 / SPIN640Plus Autoanalyzer. Accuracy: Results obtained using SPINREACT reagents (y) did not
 General laboratory equipment. show systematic differences when compared with other commercial
reagents (x).
SAMPLES The results obtained using 50 samples were the following:
 Urine 24 h: Stability 8 days at 2-8ºC. Correlation coefficient (r) 2: 0,9338
 Cerebrospinal fluid (CSF): Stable 4 days at 2-8ºC. Regression equation: y = 0,4294x – 5,4159
The results of the performance characteristics depend on the analyzer
REFERENCE VALUES5 used.
Urine: <100 mg/24h (<150 mg/24h in pregnancy)
NOTES
Children 300 -1000 mg/L
CSF: 1. In order to avoid possible contaminations, it is recommended
Adults 150 - 450 mg/L
cleaning the needle after this parameter determination. Due to its
These values are for orientation purpose; each laboratory should content the system can be easily contaminated.
establish its own reference range.
BIBLIOGRAPHY
QUALITY CONTROL 1. Orsonneau JL et al. An improved Pyrogallol Red-Molybdate Method for
A calibration using a urine protein calibrator is needed (Ref.: Determining Total Urinary Protein. Clin Chem 1989 (35):2233-2236.
2. Koller A. Total serum protein. Kaplan A et al. Clin Chem The C.V. Mosby
1002450). Each laboratory should establish its own Quality Co. St Louis. Toronto. Princeton 1984; 1316-1324 and 418.
Control scheme and corrective actions if controls do not meet the 3. Young DS. Effects of drugs on Clinical Lab. Tests, 4th ed AACC Press,
acceptable tolerances. 1995.
4. Young DS. Effects of disease on Clinical Lab. Tests, 4th ed AACC 2001.
BARCODED REAGENTS LOAD MUST BE PRECEDED OF A 5. Burtis A et al. Tietz Textbook of Clinical Chemistry, 3rd ed AACC 1999.
SPINREACT “DATABASE” COPY INTO THE ANALYZER 6. Tietz N W et al. Clinical Guide to Laboratory Tests, 3rd ed AACC 1995.
SOFTWARE. IT IS AVAILABLE UNDER REQUEST TO PACKAGING
SPINREACT.
Ref: MD1001025 Cont. R: 6 x 40 mL

MDBSIS29-I 27/04/17 SPINREACT,S.A./ S.A.U. Ctra.Santa Coloma, 7 E-17176 SANT ESTEVE DE BAS (GI) SPAIN
Tel. +34 972 69 08 00 Fax +34 972 69 00 99. e-mail: [email protected]

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