HBsAg ELISA - 3rd Gen

HBsAg ELISA - 3rd Gen

ProLISA HBsAg 3.0 ELISA is an in vitro enzyme immunosorbent assay for the qualitative determination of Hepatitis B surface antigen concentration in human serum or plasma. It is intended for professional use only as an aid in early diagnosis of Hepatitis B virus and for clinical diagnostic testing.

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Overview:

  • HBsAg 3.0 ELISA is an in vitro enzyme immunosorbent assay for the qualitative determination of Hepatitis B surface antigen concentration in human serum or plasma.

  • Caused by the Hepatitis B virus that affects the liver.

  • Causes acute and chronic infections.

  • HBsAg virus is transmitted through-

  • Sexual contact

  • Contaminated blood and blood products

  • Blood transfusion

  • Vertical transmission from mother to child during childbirth.​


 

Principle:

“Antibody sandwich technique.”

  • Recombinant anti-HBsAg specific monoclonal antibodies are coated onto microwells.

  • Samples with positive and negative controls are added in the coated wells and incubated.

  • Wash the wells to remove unbound components and monoclonal an hepatitis B monoclonal antibody conjugated to horseradish peroxidase (HRP) is added.

  • After washing wells to remove unbound enzyme, substrate is added.

  • Stop solution is added

  • Color is read on photometer at 450/650 nm wavelength.



 

Procedure:

  1. Bring all the reagents and specimen to room temperature before use.

  2. Take the required number of strips and fix them to frame and immediately close the pouch.

  3. Prepare template in data sheet indicating the location of controls and specimens.

  4. Take A1 as a Blank.

  5. Add 50µl of negative control in each well no. B1, C1 and D1 respectively.

  6. Add 50µl positive control in E1 and F1 wells.

  7. Add 50µl of specimen in each well starting from G1.

  8. Add 100µl of working enzyme conjugate to each well except Blank.

  9. Cover the plate with aluminum foil and incubate for 60 minutes at 37°C.

  10. After incubation, aspirate the contents from all the wells and wash each well 5 times with by filling approximately 300µl diluted wash buffer.

  11. Invert the plate and tap it on absorbent paper to remove the remaining washing solution, and then pipette 100µl of prepared diluted substrate into each well and incubate at room temperature for 30 minutes.

  12. Add 100µl of stop solution each well.

  13. Read absorbance at 450nm within 30 minutes in ELISA READER.


 

  • Store all kit components at 28°C when not in use.

  • Sensitivity 100% Specificity >=98%