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Journal of Clinical Microbiology, October 2009, p. 3276-3282, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.00750-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Serological Immunoassay for Detection of Hepatitis E Virus on the Basis of Genotype 3 Open Reading Frame 2 Recombinant Proteins Produced in Trichoplusia ni Larvae{triangledown}

Nereida Jiménez de Oya,1 Inmaculada Galindo,1 Olivia Gironés,2 Erwin Duizer,3 José M. Escribano,1 and Juan-Carlos Saiz1*

Department of Biotechnology, INIA, Madrid,1 Department of Infectious Pathology and Epidemiology, Facultad de Veterinaria, Universidad de Zaragoza, Saragossa, Spain,2 National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands3

Received 14 April 2009/ Returned for modification 13 July 2009/ Accepted 26 July 2009

Hepatitis E virus (HEV) is a major cause of acute hepatitis in humans, and strains of genotypes 1 and 2 are endemic in many regions with suboptimal sanitary conditions. In many industrialized countries, HEV strains of genotype 3 are highly endemic in swine, and an increased number of autochthonous infections with HEV genotype 3 strains have been reported lately. Serological studies of HEV infection are often conducted with commercial assays based on peptides and recombinant proteins of HEV genotype 1 and 2 strains. For some patients with proven HEV genotype 3 infections, these assays failed to detect specific antibodies, and they are not applicable or validated for the detection of anti-HEV antibodies in swine. To elucidate the incidence of hepatitis E in regions where HEV genotype 3 infections can be expected, and to study the seroprevalence of HEV in swine, new tools with broad specificity for all genotypes of HEV are needed. We present the expression and partial characterization of recombinant HEV genotype 3 open reading frame 2 (ORF-2) proteins and their usefulness as diagnostic antigens in detecting anti-HEV antibodies in humans and swine with proven HEV genotype 3 infections. The recombinant antigens were produced at relatively high yields and at low cost upon infection of Trichoplusia ni larvae with recombinant baculoviruses expressing recombinant HEV genotype 3 ORF-2 proteins. The enzyme-linked immunosorbent assay based on the recombinant proteins showed good specificity and sensitivity for anti-HEV genotype 3 immunoglobulin G detection in human and swine sera. These recombinant HEV genotype 3 ORF-2 proteins might be added to diagnostic kits containing HEV genotype 1 and 2 antigens in order to develop a broadly sensitive new tool for the diagnosis of hepatitis E.

* Corresponding author. Mailing address: Departamento de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Ctra. Coruña, Km. 7.5, 28040 Madrid, Spain. Phone: 34-91-3471497. Fax: 34-91-3572293. E-mail: jcsaiz{at}inia.es

{triangledown} Published ahead of print on 5 August 2009.

Journal of Clinical Microbiology, October 2009, p. 3276-3282, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.00750-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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