Hot Sale Diagnostic Reagent Human Use With 96 Test Hbc Antibody HBcAb Elisa Kit

Intended Use

Biovantion HBcAb ELISA Kit is an in vitro enzyme linked immunoassay supplied for the detection of HBcAb in human serum or plasma. It is intended for use in clinical laboratories for diagnosis and management of patients related to infection with hepatitis B virus.

Summary

Hepatitis B is an infectious illness caused by hepatitis B virus (HBV) which is an enveloped, double-stranded DNA virus belonging to the Hepadnaviridae family and is recognized as the major cause of blood transmitted hepatitis together with hepatitis C virus (HCV). HBV is excreted in body fluids such as semen, saliva, blood and urine in persons with acute or chronic infection. The hepatitis B virus is known as a blood-borne virus because it is transmitted from one person to another via blood or fluids contaminated with blood. Transmission of hepatitis B virus results from exposure to infectious blood or body fluids.When HBV invades the body it causes liver damage through induction of auto-immunity. Three immunity has been found, surface antigen (HBsAg)/HBsAb, core antigen (HBcAg)/HBcAb and e antigen(HBeAg)/HBeAb. As it is difficult to detect the core antigen in the serum, the other five are been done to diagnosing HBV. Hepatitis B “core ” antigen (HBcAg) is a major component of the viral structure. Antibodies to HBcAg (anti-HBc total antibody, and IgM) appear shortly after the appearance of HBsAg and persist for life both in persons who have recovered from a hepatitis B infection and in those who develop HBsAg-carrier status but in rare cases, a HBV infection can also run its course without the appearance of immunologically detectable anti-HBc (usually in immunosuppressed patients). Anti-HBc is a marker of acute, chronic or resolved HBV infection and screening for anti-HBc provides with information on the prevalence of the disease in different groups. In the absence of other hepatitis B markers (HBsAg-negative persons), anti-HBc may be the only indication of an existing hepatitis B viral infection.

MATERIALS AND COMPONENTS

MICROPLATE: Blank microwell strips fixed on white strip holder. The plate is sealed in aluminum pouch with desiccant. Each well contains purified HBcAg. 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.

NEGATIVE CONTROL: (1x1.0ml per vial) preserv.0.1% ProClinTM 300 Yellowish liquid filled in a vial with green screw cap.Protein-stabilized buffer tested non reactive for HBcAb. Ready to use as supplied. Once open, stable for one month at 2-8°C.

POSITIVE CONTROL: (1x1.0ml per vial) preserv.0.1% ProClinTM 300 Red-colored liquid filled in a vial with red screw cap. HBcAb diluted in protein-stabilized buffer. 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 monoclonal antibodies to HBcAg. 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 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 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.

PROCEDURE

Reagents preparation: Allow the reagents to reach room temperature (18-30°C). Dilute the Wash buffer (20X) as indicated in the instructions for washing. Use distilled or deionized water and only clean vessels to dilute the buffer. All other reagents are READY TO USE AS SUPPLIED.

1. Preparation: Format the microplate’s wells for control and patient specimen to be assayed. Replace any unused microwell strips back into the aluminum bag seal and store at 2-8°C.
2. Adding Sample: Add 50μl of the control or the samples into the assigned well.
3. Adding Conjugate: Add 50μl of CONJUGATE into each well except the Blank.
4. Incubating: Cover the plate with the plate cover and incubate for 30 minutes at 37°C.
5. Washing: At the end of the incubation, remove and discard the plate cover. Wash each well 5 times with diluted Wash Buffer. Each time allow the microwells to soak for 30-60 seconds. After the final washing cycle, turn down the plate onto blotting paper or clean towel and tap it to remove any remainders.
6. Adding Substrate: Add 50μl of Substrate Solution A and 50μl of Substrate Solution B into each well. Incubate for 10 minutes at 37°C avoiding light.
7. Adding Stop Solution: Using a multichannel pipette or manually, add 50μl of Stop Solution into each well and mix gently.
8. Measuring the Absorbance: Calibrate the plate reader with the Blank well and read the absorbance at 450nm. If a dual filter instrument is used, set the reference wavelength at 630nm. Calculate the Cut-off value and evaluate the results. (Note: read the absorbance within 10 minutes after stopping the reaction).