Expression of non-conserved regions of the S genome segments of three hantaviruses: evaluation of the expressed polypeptides for diagnosis of haemorrhagic fever with renal syndrome

Abstract

Haemorrhagic fever with renal syndrome (HFRS) is a serious and often fatal disease caused by viruses in the Hantavirus genus of the family Bunyaviridae. We expressed the entire coding region of the small (S) genome segments of three serologically distinct hantaviruses as soluble proteins in Escherichia coli and evaluated the expressed nucleocapsid proteins (NPs) as antigens for diagnosis of HFRS. We also prepared novel diagnostic antigens by expressing truncated genes from which we deleted amino acid coding regions that were highly conserved among the three viruses. These antigens were analysed for their potential to detect and differentiate between antisera to various hantaviruses by ELISA. ELISA results obtained with HFRS patient sera or with sera from naturally or experimentally infected animals indicate that homologous antigens and antisera reacted to high titre. The truncated NPs were more specific than the complete NPs in distinguishing between possible aetiological agents of HFRS. Our findings demonstrate that prokaryotic expression of portions of the NPs of specific hantaviruses can be used to generate, readily and efficiently, large quantities of antigen that is both sensitive and specific in diagnostic assays for HFRS.

Development and evaluation of a solid-phase enzyme immunoassay based on Andes hantavirus recombinant nucleoprotein

“…The association of hantaviruses with different rodent reservoirs in several geographic areas suggests that development of region-specific antigens should be undertaken to improve serological reactivity. As for other viruses, the use of the homologous antigen improves the sensitivity and specificity of the assays, providing earlier antibody detection in infected people [8][9][10][11]. Thus, some of the South American viruses may give higher optical density readings in enzyme-linked immunosorbent assays (ELISA) with reagents based on the local viral sequence [12].…”

A soluble and heat-resistant form of hantavirus nucleocapsid protein for the serodiagnosis of HFRS

“…In addition, it possesses immunodominant, linear, cross-reactive epitopes in the N-terminal region (Elgh et al, 1996b;Gott et al, 1997;Jenison et al, 1994;Wang et al, 1993;Yamada et al, 1995). In order to provide large quantities of pure antigen for diagnostic purposes, the ATS peptide was introduced into N-terminal parts of HTNV-N and SEOV-N (1-70 amino acids residues of N) by recombinant DNA technology (Fig.…”

“…Due to the practical limitations of IFA, many researchers have made efforts to use an ELISA method for the serodiagnosis of HFRS. In particular, recombinant viral protein is used to eliminate the risks associated with processing of native antigens and to produce high quality antigens for ELISA (Elgh et al, 1995;Kallio-Kokko et al, 1993;Vapalahti et al, 1996;Wang et al, 1993;Zoller et al, 1993). However, the viral protein used for ELISA is usually expressed as inclusion bodies in E. coli, and requires an in vitro refolding process before purification .…”

“…One is located within the N-terminal 1-120 amino acids and contains linear and cross-reactive epitopes to other hantaviruses (Elgh et al, 1996b;Gott et al, 1997;Jenison et al, 1994;Wang et al, 1993;Yamada et al, 1995). The other is located within the C-terminus and contains conformation-dependent and serotypespecific epitopes (Araki et al, 2001;Morii et al, 1998;Ruo et al, 1991;Yoshimatsu et al, 1996).…”

Expression of the Nucleoprotein of the Puumala Virus from the Recombinant Semliki Forest Virus Replicon: Characterization and Use as a Potential Diagnostic Tool

“…In addition, some hantaviruses must be manipulated in a high-security containment facility. Therefore, several laboratories have expressed the N protein as a recombinant protein in Escherichia coli (6,9,24,37) or in insect cells (5,29,30,33,36).…”