Serodiagnosis using recombinant nipah virus nucleocapsid protein expressed in Escherichia coli.
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.
Yu
Fuxun
F
Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Khairullah
Nor Shahidah
NS
Inoue
Shingo
S
Balasubramaniam
Vijayamalar
V
Berendam
Stella Joan
SJ
Teh
Leok Kin
LK
Ibrahim
Nik Shamsiah Wan
NS
Abdul Rahman
Sohayati
S
Hassan
Sharifah Syed
SS
Hasebe
Futoshi
F
Sinniah
Mangalam
M
Morita
Kouichi
K
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D045464:Henipavirus Infections / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D045405:Nipah Virus / Q000276:immunology*; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000276:immunology / Q000821:virology
D000914:Antibodies, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins