Serodiagnosis using recombinant Nipah virus nucleocapsid protein expressed in Escherichia coli

Fuxun Yu, Nor Shahidah Khairullah, Shingo Inoue, Vijayamalar Balasubramaniam, Stella Joan Berendam, Leok Kin Teh, Nik Shamsiah Wan Ibrahim, Sohayati Abdul Rahman, Sharifah Syed Hassan, Futoshi Hasebe, Mangalam Sinniah, Kouichi Morita

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34 Citations (Scopus)

Abstract

Nipah virus nucleocapsid (NiV-N) protein was expressed in Escherichia coli and purified by histidine tag-based affinity chromatography. An indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) for human and swine sera and an IgM capture ELISA for human sera were established using the recombinant NiV-N protein as an antigen. One hundred thirty-three suspected patient sera and 16 swine sera were used to evaluate the newly established ELISA systems in comparison with the CDC inactivated-virus-based ELISA systems. For the human sera, the NiV-N protein-based indirect IgG ELISA had a sensitivity of 98.6% and a specificity of 98.4%, and the NiV-N protein-based IgM capture ELISA had a sensitivity of 91.7% and a specificity of 91.8%, with reference to the CDC ELISA systems. The NiV-N-based IgM ELISA was found to be more sensitive than the inactivated-virus-based ELISA in that it captured eight additional cases. For the swine sera, the two test systems were in 100% concordance. Our data indicate that the Nipah virus nucleocapsid protein is a highly immunogenic protein in human and swine infections and a good target for serodiagnosis. Our NiV-N protein-based ELISA systems are useful, safe, and affordable tools for diagnosis of Nipah virus infection and are especially fit to be used in large-scale epidemiological investigations and to be applied in developing countries.

Original languageEnglish
Pages (from-to)3134-3138
Number of pages5
JournalJournal of Clinical Microbiology
Volume44
Issue number9
DOIs
Publication statusPublished - Sep 2006
Externally publishedYes

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