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Clin. Vaccine Immunol. doi:10.1128/CVI.00186-07
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

A Double-Antigen Enzyme-Linked Immunosorbent Assay for Detection of Antibodies to Hepatitis E Virus in Human or Swine Sera

MP Biomedicals Asia Pacific Pte Ltd. (formerly Genelabs Diagnostics Pte Ltd), Singapore; Macfarlane Burnet Institute for Medical Research and Public Health, Melbourne, Australia; and Select Vaccines Ltd, Melbourne, Australia

^* To whom correspondence should be addressed. Email: ming.guan{at}mpbio.com.

	Abstract

A new double-antigen sandwich-based enzyme-linked immunosorbent assay for the detection of total antibodies (IgG and IgM) specific to hepatitis E virus was developed utilizing the well-characterized recombinant protein ET2.1 and its peroxidase-labeled counterpart. Our study showed that the ELISA detected all the positive patient samples (n=265) regardless of whether they contained IgM or IgG antibodies or both, while maintaining an excellent specificity of 98.8% with samples from various patient or healthy control groups (Total n=424). The test had a detection limit for anti-HEV IgG antibodies at a 62mIU/ml equivalent of the international reference. When compared with the serological status of the specimens based on testing at the individual collection sites, the testing outcome generated by the new ELISA had a good agreement of 99.3% with a kappa value of 0.985. The positive predictive value (PPV) and the negative predictive value (NPV) for the new test reached 98.1% and 100%, respectively. This ELISA had a positive delta of 4.836 and a negative delta of 3.314 indicating an excellent differentiation of the infected and non-infected cohorts. Furthermore, the new design enables the detection of antibodies not only in human but also in pig samples. Our preliminary data showed that the ELISA could detect seroconversion samples from pigs as early as 14 days post inoculation. The potential utility of detecting specific antibodies in pigs will be an added advantage for managing the disease with suggested zoonotic implications.