RPR Carbon

RPR Carbon - The RPR-carbon is a non-treponemal slide agglutination test for the qualitative and semi-quantitative detection of plasmatic reagins in human serum. Carbon particles coated with a lipid complex are agglutinated when mixed with samples containing reagins. The reagent sensitivity is calibrated against the “Human Reactive Serum” from CDC (Centre for Disease Control). Diagnostic sensitivity: 100 % Diagnostic specificity: 100 % REAGENTS COMPOSITION: 2,5 mL Carbon: Carbon particles coated with a lipid complex, cardiolipin, lecithin and cholesterol, in phosphate buffer 20 mmol/L. Sodium azide 0.95 g/L. pH, 7.0 1 mL Control (+): Human serum with a reagin titer ≥ 1/4. Sodium azide 0.95 g/L. 1 mL Control (-): Animal serum. Sodium azide 0.95 g/L. 8X6 slids MATERIAL REQUIRED BUT NOT PROVIDED: Mechanical rotator with adjustable speed at 80-100 r.p.m. PRECAUTIONS: Components from human origin have been tested and found to be negative for the presence of HBsAg and HCV, and of antibody to HIV (1/2). However handle cautiously as potentially infectious. Good laboratory safety practices should be followed when handling laboratory reagents or human samples. TEST PROCEDURE: Preparation RPR-carbon: Shake the reagent gently to disperse the carbon particles before use. Open the RPR-carbon vial, place the micropipette to the dispensing vial and draw by suction the required volume of RPR-carbon. Once the test is completed, return the reagent to the original vial and rinse the micropipette and vial with distilled water. Qualitative method 1. Allow the reagents and sample to reach room temperature. The sensitivity of the test may be reduced at low temperatures. 2. Place 50 μL of the sample and one drop of each Positive and Negative control into separate circles on the slide test. 3. Shake the RPR-carbon reagent gently before using. Invert the dropper assembly and press gently to remove air bubbles from the micropipette. 4. Place the micropipette in a vertical position and perpendicular to the slide, and add a drop of this reagent next to the sample to be tested. 5. Mix both drops with a stirrer, spreading them over the entire surface of the circle. Use different stirrers for each sample. 6. Rotate the slide with a mechanical rotator at 80-100 r.p.m. for 8 minutes. False positive results could appear if the test is read later than 8 minutes. Semi-quantitative method 1. Make serial two fold dilutions of the sample in 9 g/L saline solution. 2. Proceed for each dilution as in the qualitative method. READING AND INTERPRETATION: Examine macroscopically the presence or absence of visible agglutination immediately after removing the slide test from the rotator. Rotate the slide twice by hand before reading. The titer, in the semi-quantitative method, is defined as the highest dilution showing a positive result. QUALITY CONTROL: Positive and Negative controls are recommended to monitor the performance of the procedure, as well as a comparative pattern for a better result interpretation. Serum controls RF are recommended for internal quality control. Each laboratory should establish its own Quality Control scheme and corrective actions. CLINICAL SIGNIFICANCE: Reagins are a group of antibodies against some components of the damage tissues from patients infected by Treponema pallidum, the agent which causes the syphilis .This microorganism produces some damage to the liver and heart, releasing some tissue fragments. Immunological patient system reacts producing regains, -antibodies against these fragments.