Abstract
The early diagnosis of West Nile virus (WNV) infection is important for successful clinical management and epidemiological control. The non-structural protein 1 (NS1) of flavivirus, a highly conserved and secreted glycoprotein, is abundant in the serum of flavivirus-infected patients and represents a useful early diagnostic marker. We developed a WNV-specific NS1 antigen-capture ELISA using two mouse monoclonal antibodies (MAbs) that recognised distinct epitopes of the NS1 protein of WNV as capture and detection antibodies. The antigen-capture ELISA displayed exclusive specificity to WNV without cross-reaction with other related members of the flavivirus family, including the dengue virus, yellow fever virus, Japanese encephalitis virus, and tick-borne encephalitis virus. Additionally, the specificity was presented as no false positive in normal (0/1003) and DENV-infected (0/107) human serum specimens. The detection limit of the antigen-capture ELISA was as low as 15 pg/ml of recombinant WNV NS1 protein (rWNV-NS1) and 6.1 plaque-forming units (PFU)/0.1 ml of WNV-infected culture supernatant. In mice infected with WNV, the NS1 protein was readily detected in serum as early as one day after WNV infection, prior to the development of clinical signs of the disease. The sensitivity of the NS1 capture ELISA (93.7%) was significantly higher (79.4%) than that of real-time reverse transcription polymerase chain reaction in 63 serum samples from WNV-infected mice (p = 0.035). This newly developed NS1 antigen-capture ELISA with high sensitivity and specificity could be used as an efficient method for the early diagnosis of WNV infection in animals or humans.
Highlights
West Nile virus (WNV), a member of the flavivirus family, is closely related to significant human pathogens such as yellow fever virus (YFV), dengue virus (DENV) type 1 to 4, Japanese encephalitis virus (JEV), tick-borne encephalitis virus (TBEV), St
A total of 65 hybridoma cell clones that produced monoclonal antibodies (MAbs) against WNV-non-structural protein 1 (NS1) protein were established based on their strong positive reactivity with the rWNV-NS1 protein in the ELISA and the WNV-infected cells in the immunofluorescence assay (IFA)
The specificity of the MAbs was further identified by excluding their cross-reactivity to the four serotypes of DENV, JEV, and YFV using IFA and the four serotypes of DENV-NS1 protein using ELISA
Summary
West Nile virus (WNV), a member of the flavivirus family, is closely related to significant human pathogens such as yellow fever virus (YFV), dengue virus (DENV) type 1 to 4, Japanese encephalitis virus (JEV), tick-borne encephalitis virus (TBEV), St. WNV is transmitted through a mosquito-birdmosquito route and causes a range of illnesses in humans including mild fever, acute flaccid paralysis, and lethal encephalitis. WNV presents a global threat to human and animal health across the United States and Canada as well as in temperate regions such as Europe and Africa [1]. The public health concern regarding these diseases attracted increasing attention during the last decade in WNV endemic areas of Europe and the Middle East [2]. Since 1999, WNV has spread rapidly throughout North America, causing epidemics of WNV encephalitis, meningitis, and acute flaccid paralysis in naive populations. There were 39,462 cases of WNV infection in the United States from 1999 to 2013, and 1663 deaths have been reported (http://www.cdc.gov/ncidod/dvbid/ westnile/index.htm)
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