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Clinical and Vaccine Immunology, August 2008, p. 1151-1157, Vol. 15, No. 8
1071-412X/08/$08.00+0     doi:10.1128/CVI.00186-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Double-Antigen Enzyme-Linked Immunosorbent Assay for Detection of Hepatitis E Virus-Specific Antibodies in Human or Swine Sera {triangledown}

Wei Ping Hu,1 Yang Lu,1 Nestor Amadeo Precioso,1 Hsiao Ying Chen,1 Teresa Howard,2,3 David Anderson,2,3 and Ming Guan1*

MP Biomedicals Asia Pacific Pte Ltd., Singapore, Republic of Singapore,1 Macfarlane Burnet Institute for Medical Research and Public Health, Melbourne, Australia,2 Select Vaccines Ltd., Melbourne, Australia3

Received 4 May 2007/ Returned for modification 3 October 2007/ Accepted 14 May 2008

A new double-antigen sandwich-based enzyme-linked immunosorbent assay (ELISA) for the detection of total antibodies (immunoglobulin G [IgG] and IgM) specific for hepatitis E virus (HEV) was developed by utilizing 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 it maintained an excellent specificity of 98.8% with samples from various patient or healthy control groups (total number of samples, 424). The test had a detection limit for anti-HEV IgG antibodies that was equivalent to 62 mIU/ml of the international reference. Compared with the serological status of the specimens determined on the basis of tests performed 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 and the negative predictive value for the new test reached 98.1% and 100%, respectively. This ELISA had a positive delta value of 4.836 and a negative delta value of 3.314 (where delta is a measure of the number of standard deviations by which the cutoff is separated from the mean of the sample groups) (N. Crofts, W. Maskill, and I. D. Gust, J. Virol. Methods 22:51-59, 1988), indicating that it had an excellent ability to differentiate the infected and noninfected cohorts. Furthermore, the new design enables the detection of antibodies not only in human samples but also in pig samples. Our preliminary data showed that the ELISA could detect seroconversion in samples from pigs at as early as 14 days postinoculation. The potential utility of detecting specific antibodies in pigs will be an added advantage for managing the disease, with suggested zoonotic implications.

* Corresponding author. Present address: BD Rapid Diagnostics (Suzhou) Ltd., 9 Rui Pu Road, Export Processing Zone B, Suzhou Industrial Park, Jiangsu, People's Republic of China. Phone: 86-512-67632708. Fax: 86-512-67632898. E-mail: guan_ming{at}bd.com

{triangledown} Published ahead of print on 21 May 2008.

Clinical and Vaccine Immunology, August 2008, p. 1151-1157, Vol. 15, No. 8
1071-412X/08/$08.00+0     doi:10.1128/CVI.00186-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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