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Am. J. Trop. Med. Hyg., 48(3), 1993, pp. 440-446
Copyright © 1993 by The American Society of Tropical Medicine and Hygiene

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Detection of West Nile Virus by the Polymerase Chain Reaction and Analysis of Nucleotide Sequence Variation

Kevin R. Porter, Peter L. Summers, Doria Dubois, Beena Puri, William Nelson, Erik Henchal, John J. Oprandy AND Curtis G. Hayes
Infectious Diseases Threat Assessment Program, Naval Medical Research Institute, Bethesda, Maryland; Walter Reed Army Institute of Research, Washington, District of Columbia

A polymerase chain reaction (PCR) assay was developed to rapidly detect and identify West Nile (WN) virus. The RNA from seven isolates of WN virus from six countries and four other flaviviruses (Kunjin, Japanese encephalitis, St. Louis encephalitis, and yellow fever viruses) was reverse-transcribed (RT) and amplified by PCR. The nucleotide sequences of the amplified products were determined by a rapid, automated DNA sequencing method. The WN virus RT/PCR assay detected the target gene segment of isolates from both the African-Middle Eastern group and the Indian group with a sensitivity of approximately 0.05 pg of viral RNA. Kunjin virus was the only other flavivirus tested that produced a band of the appropriate size. Five of seven WN virus isolates showed 92–98% homology in the nucleotide sequence of their PCR products. The sequence of one isolate was virtually identical to the published sequence of the Nigerian isolate (99.5% homology). No correlation was established between the degree of nucleotide homology, geographic location, time of isolation, or source of the isolates.




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V. P. Bondre, R. S. Jadi, A. C. Mishra, P. N. Yergolkar, and V. A. Arankalle
West Nile virus isolates from India: evidence for a distinct genetic lineage
J. Gen. Virol., March 1, 2007; 88(3): 875 - 884.
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Copyright © 1993 by the American Society of Tropical Medicine and Hygiene.