Improving Access to Anti-HDV Testing: Development and Validation of an Affordable In-House ELISA Assay

Overview

Journal ACS Omega

Publisher American Chemical Society

Specialty Chemistry

Date 2024 Apr 22

PMID 38645367

Authors

Iram Amin

Shazia Rafique

Amjad Ali

Nadeem Ahmed

Muhammad Shahid

Samia Afzal

Saad Tahir

Muhammad Waqas

Sadia Bibi

Abdallah M Elgorban

Muhammad Idrees

Masaud Shah

Asad Syed

Affiliations

Soon will be listed here.

Abstract

In certain low-income nations, the hepatitis Delta virus and hepatitis B virus (HBV) pose a serious medical burden, where the prevalence of hepatitis B surface antigen (HBsAg) is greater than 8%. Especially in rural places, irregular diagnostic exams are the main restriction and reason for underestimation. Utilizing serum samples from a Pakistani isolate, an internal ELISA for the quick identification of anti-HDV was created, and the effectiveness of the test was compared to a commercial diagnostic kit. HDV-positive serum samples were collected, and a highly antigenic domain of HDAg antigen was derived from them. This antigenic HDAg was expressed in a bacterial expression system, purified by Ni-chromatography, and confirmed by SDS-PAGE and Western blot analysis. The purified antigen was utilized to develop an in-house ELISA assay for anti-HDV antibody detection of the patient's serum samples at very low cost. Purified antigens and positive and negative controls can detect anti-HDV (antibodies) in ELISA plates. The in-house developed kit's efficiency was compared with that of a commercial kit (Witech Inc., USA) by the mean optical density values of both kits. No significant difference was observed (a value of 0.576) by applying statistical analysis. The newly developed in-house ELISA is equally efficient compared to commercial kits, and these may be useful in regular diagnostic laboratories, especially for analyzing local isolates.

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