Comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivat
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RESEARCH
Open Access
Comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivated virus as antigens Fuxun Yu1 , Ferdinard Adungo2,3, Samson Limbaso Konongoi3, Shingo Inoue2, Rosemary Sang3, Salame Ashur3, Allan ole Kwallah3, Leo Uchida2, Corazon C Buerano2, Matilu Mwau3, Yan Zha1, Yingjie Nie1* and Kouichi Morita2*
Abstract Background: Rift Valley Fever (RVF) is a mosquito-borne viral zoonosis. To detect RVF virus (RVFV) infection, indirect immunoglobulin G (IgG) and immunoglobulin M (IgM) enzyme linked immunosorbent assays (ELISAs) which utilize recombinant RVFV nucleocapsid (RVFV-N) protein as assay antigen, have reportedly been used, however, there is still a need to develop more sensitive and specific methods of detection. Methods: RVFV-N protein was expressed in Escherichia coli (E. coli) and purified by histidine-tag based affinity chromatography. This recombinant RVFV-N (rRVFV-N) protein was then used as antigen to develop an IgG sandwich ELISA and IgM capture ELISAs for human sera. Ninety six serum samples collected from healthy volunteers during the RVF surveillance programme in Kenya in 2013, and 93 serum samples collected from RVF-suspected patients during the 2006–2007 RVF outbreak in Kenya were used respectively, to evaluate the newly established rRVFV-N protein-based IgG sandwich ELISA and IgM capture ELISA systems in comparison with the inactivated virus-based ELISA systems. Results: rRVFV-N protein-based-IgG sandwich ELISA and IgM capture ELISA for human sera were established. Both the new ELISA systems were in 100% concordance with the inactivated virus-based ELISA systems, with a sensitivity and specificity of 100%. Conclusions: Recombinant RVFV-N is a safe and affordable antigen for RVF diagnosis. Our rRVFV-N-based ELISA systems are safe and reliable tools for diagnosis of RVFV infection in humans and especially useful in large-scale epidemiological investigation and for application in developing countries. Keywords: RVFV, Nucleocapsid protein, Expression, IgG sandwich ELISA, IgM capture ELISA
Background Rift Valley Fever (RVF) is a mosquito-borne viral zoonosis, which periodically causes disease outbreaks in humans and livestock. Infection by RVF virus (RVFV) causes abortion or resorption of the fetus in pregnant domestic ruminants, with newborn mortality approaching 100%. Transmission of RVFV to humans, either through contact with bodily * Correspondence: [email protected]; [email protected] 1 Guizhou Provincial People’s Hospital, Medical College, Guizhou University, No. 83 Zhongshan Dong Road, Guiyang 550002, Guizhou Province, China 2 Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12- 4, Sakamoto, Nagasaki 852-8523, Japan Full list of author information is available at the end of the article
fluids of infected animals or through mosquito bites, may result in mild to moderate influenza-like symptoms and severe retinitis, encephalitis and hemorrhagic fever [1, 2]. RVFV was first described i
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