HBsAb Quantitative ELISA Test Kit

INTENDED USE

  The purpose of the HBsAb quantitative ELISA Test is for in vitro quantitative determination of antibodies to hepatitis B virus surface antigen in human serum or plasma for clinical purposes and assessing antibody response levels to HBsAg vaccine.

PRINCIPLE OF THE TEST

For HBsAb quantitative ELISA test kit uses antigen “sandwich” ELISA method in which, polystyrene microwell strips are pre-coated with recombinant HBsAg. The test begins by having a combination of patient's serum or plasma sample being added to Horseradish Perioxidase (HRP-Conjugate) to the microwells. If anti-HBs is present, the pre-coated and conjugated antigens will be bound to the two variable domains of the antibody. A specific immunocomplex is formed and captured on the solid phase during incubation. When washing is complete (to remove unbound HRP-conjugates and sample serum proteins), chromogen solutions containing Tetramethylbenzidine (TMB) and urea peroxide are added to the wells. The colorless chromogens are hydrolyzed by the bound HRP-Conjugate to a blue-colored product when there is a presence of antigen-antibody-antigen (HRP) sandwich complex. After stopping the reaction with sulfuric acid, the blue color turns yellow. The color intensity can be gauged proportionally to the amount of antibody captured in the wells, and to the amount in the sample, respectively. The wells remain colorless if the anti-HBs result is negative.

MATERIALS AND COMPONENTS

  This kit contains reagents sufficient for testing of maximum of 91 specimens in a test run. MICROPLATE: Blank microwell strips fixed on white strip holder. The plate is sealed in aluminum pouch with desiccant. Each well contains recombinant HBsAg. The microwell strips can be broken to be used separately. Place unused wells or strips in the provided plastic sealable storage bag together with the desiccant and return to 2-8°C. Once open, stable for one month at 2-8°C. CALIBRATORS:(1x0.5ml per vial) Six vials of references for HBsAb at levels of 0(A),10.0(B),20.0(C),40.0(D),80.0(E) and 160.0(F) mIU/ml. Store at 2-8°C.Ready to use as supplied. Once open, stable for one month at 2-8°C. CONJUGATE: (1x6ml per vial) preserv.0.1% ProClinTM 300 Red-colored liquid in a white vial with red screw cap. Horseradish peroxidase-conjugated HBsAg antigens. Ready to use as supplied. Once open, stable for one month at 2-8°C. WASH BUFFER: (1x20ml per bottle) DILUTE BEFORE USE! detergent Tween-20 Colorless liquid filled in a white bottle with white screw cap, PH 7.4, 20 × PBS. The concentrate must be diluted 1 to 19 with distilled/deionized water before use. Once diluted, stable for one week at room temperature, or for two weeks when stored at 2-8°C. SUBSTRATE SOLUTION A: (1x6ml per vial) Colorless liquid filled in a white vial with green screw cap. Urea peroxide solution. Ready to use as supplied. Once open, stable for one month at 2-8°C. SUBSTRATE SOLUTION B: (1x6ml per vial) Colorless liquid filled in a black vial with black screw cap. TMB (Tetramethyl benzidine) solution. Ready to use as supplied. Once open, stable for one month at 2-8°C. STOP SOLUTION: (1x6ml per vial) Colorless liquid in a white vial with white screw cap. Diluted sulfuric acid solution (0.5M H2SO4).Ready to use as supplied. Once open, stable for one month at 2-8°C. PLASTIC SEALABLE BAG: For enclosing the strips not in use 1unit PACKAGE INSERT 1copy CARDBOARD PLATE COVER 2sheets To cover the plates during incubation and prevent evaporation or contamination of the wells.

MATERIALS REQUIRED BUT NOT PROVIDED

  Freshly distilled or deionized water, disposable gloves and timer, appropriate waste containers for potentially contaminated materials, dispensing system and/or pipette, disposable pipette tips, absorbent tissue or clean towel, dry incubator or water bath, 37±0.5°C, plate reader, single wavelength 450nm or dual wavelength 450/630nm, microwell aspiration/wash system.