Advantage
1.Certificate:ISO13485:2003, GMP
2.Sensitivity>9/10
3.Specificity>14/15
4.Precision: CV<15%
INTEND USED
Anti-HIV1+2 is an enzyme-linked immunosorbent assay (ELISA) for the qualitative determination of antibodies to human immunodeficiency virus type 1 and/or 2 in human serum or plasma.
KIT COMPONENTS
- Microplate 1 plate (96 wells). 12 strips per plate, each with 8 wells coated with a mixture of HIV-1 gp120, gp41, group O antigen and HIV-2 gp36.
- Positive Control 1 vial (0.5ml). Inactivated Human serum containing antibodies to HIV antigens. Preservative: Bronidox (2ml/L).
- Negative Control 1 vial (0.5ml). Inactivated Human serum without antibodies to HIV antigens. Preservative: Bronidox (2ml/L).
- Conjugate 1 vial (6ml). The solution contains HRP-labeled HIV-1 gp120, gp41, group O antigen and HIV-2 gp36, with phosphate buffer (0.02mol/L) and stabilizing proteins, ready to use.
- Sample Diluent 1 vial (6ml). Contains stabilizing proteins and detergent, Preservative: Bronidox (2ml/L).
- Wash Buffer (25´) 1 vial (30ml). Diluted 25-fold in distilled water as described in section of Preparation of Reagent.
- Substrate Solution A 1 vial (6ml). Hydrogen peroxide-urea (approx. 0.6g/L) in citrate-acetate buffer solution (10 mmol/L).
- Substrate Solution B 1 vial (6ml). Tetramethyl benzidine dihydrochloride (10g/L).
- Stop Solution 1 vial (6ml). 2N sulphuric acid.
- Plate Covers 4 pieces
- Instruction Manual 1 copy
ASSAY PROCEDURE
- Add 100ml of sample diluent to wells A1 and B1as blank control.
- Dispense 50ml of sample diluent into the other wells.
- Add 50ml of negative control to well C1 and D1 and add 50ml of positive control to wells E1 and F1.
- Add 50ml of specimen to the assigned well , starting at well G1.
- Carefully cover microplate with a plate cover provided to prevent evaporation during incubation.
- Incubated for 45 minutes at 37°C.
- Wash the microplate with diluted wash buffer.
A.ELISA Microplate Washer-Wash 5 times with at least 300 ml/well/wash.
B.Manual Microplate Washer-Aspirate completely the contents of all wells by lowering the aspirator tip gently to the bottom of each well. Be careful not to scratch the inside of the well surface. Fill the entire plate with at least 300 ml/well, then aspirate in the same order. Perform this cycle 5 times.
- Blot dry by inverting the microplate and tapping firmly onto absorbent paper. All residual plate wash buffer should be blotted dry.
- Add 50ml of conjugate to each well (except for the blank control), apply another plate cover.
- Incubate for 30 minutes at 37°C.
- Repeat the wash procedure as in Step 7 and Step 8.
- Add 50ml of substrate solution A and 50ml substrate solution B to each well.
- Incubate for 10 minutes at 37°C , away from direct or intense light .
- Stop the reaction by adding 50ml of stop solution to each well.
- Determine the absorbance for each well at 450nm against a reference of 630nm with a microplate reader.
