Development and evaluation of an in-house ELISA to detect hepatitis B virus surface antigen in resource-limited settings

Authors

  • K Fatema Department of Pathology & Microbiology, City Dental College, Malibagh Chowdhury Para, Dhaka
  • S Tabassum Department of Virology, Bangabandhu Sheikh Mujib Medical University, Shahbag, Dhaka
  • A Nessa Department of Virology, Bangabandhu Sheikh Mujib Medical University, Shahbag, Dhaka
  • M Jahan Department of Virology, Bangabandhu Sheikh Mujib Medical University, Shahbag, Dhaka

DOI:

https://doi.org/10.3329/bmrcb.v39i2.19644

Abstract

Hepatitis B virus (HBV) infection is of global public health concern. Among various serological tests used for the diagnosis and screening of HBV infection, the enzyme-linked immunosorbent assay (ELISA) to detect hepatitis B surface antigen (HbsAg) is most widely used. The present study was designed to develop and standardize a cost effective in-house ELISA for the detection of HbsAg and compare its performance with two established commercial kits. The concentrations of coating antibody, conjugates and sera were fixed by checkerboard titration. Using known HBsAg positive and negative sera, four different concentrations (1, 0.5, 0.25 and 0.125 ?g/well) of coating anti-HBs were applied. Similarly, serial dilutions of patients sera (1 in 2, 1 in 3, 1 in 5 and 1 in 9) and conjugates (1 in 2, 1 in 3, 1 in 5, 1 in 9 and 1 in 17) were evaluated by checkerboard titration. The optimal concentration of coating antibody was determined at 0.25 ?g/well and 1 in 9 dilution for both conjugates and sera. The performance comparison of our in-house ELISA showed excellent correlation with two commercial kits (Pearson 0.957, P=0.001 for monoclonal antibody coated kit and Pearson 0.929, P=0.000 for polyclonal antibody coated kit) when OD values were compared. All commercial kit proven positive samples was positive while all negative samples were negative with the in-house ELISA resulting in 100% sensitivity and specificity. The results of our study demonstrated that our inhouse ELISA for detection of HBsAg was equally as sensitive and specific as two well-known commercial kits. Thus, this system may be a useful tool for diagnostic and screening purposes, as well as outbreak investigations.

DOI: http://dx.doi.org/10.3329/bmrcb.v39i2.19644

Bangladesh Med Res Counc Bull 2013; 39: 65-68

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Published

2014-07-23

How to Cite

Fatema, K., Tabassum, S., Nessa, A., & Jahan, M. (2014). Development and evaluation of an in-house ELISA to detect hepatitis B virus surface antigen in resource-limited settings. Bangladesh Medical Research Council Bulletin, 39(2), 65–68. https://doi.org/10.3329/bmrcb.v39i2.19644

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Section

Research Papers