Denitive identication of louping ill virus by RT-PCR and sequencing in eld populations of I.pdf

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Denitive identication of louping ill virus by RT-PCR and sequencing in eld populations of I.pdf

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Denitive identication of louping ill virus by RT-PCR and sequencing in eld populations of I

Arch Virol (1997) 142: 1181±1191 De?nitive identi?cation of louping ill virus by RT-PCR and sequencing in ?eld populations of Ixodes ricinus on the Lochindorb Estate M. W. Gaunt1, L. D. Jones1, K. Laurenson2, P. J. Hudson2, H. W. Reid3, and E. A. Gould1 1NERC, Institute of Virology and Environmental Microbiology, Oxford, U.K. 2 Wildlife Epidemiology Unit, Department of Biological and Molecular Science, University of Stirling, Stirling, U.K. 3 Moredun Research Institute, Edinburgh, U.K. Accepted January 6, 1997 Summary. Rapid and precise virus detection procedures are an important component of any epizootiological study. An automated one tube reverse transcriptase and nested primer polymerase chain reaction (RT-PCR) followed by nucleotide sequencing of the cDNA product, was used for the rapid detection and identi?cation of louping ill (LI) virus in ?eld caught Ixodes ricinus and compared with a classical isolation method i.e. infectivity in cell culture. The results establish the genetic identity of LI virus on the Lochindorb Estate. There was a high correlation between the results obtained by RT-PCR and infectivity assays. RT-PCR and sequencing proved to be a rapid and accurate system for identifying LI virus in ?eld specimens. Development of this system should improve the capacity to undertake detailed epizootiological studies of LI virus. Introduction Louping ill (LI) is a debilitating virus disease producing high rates of mortality in infected grouse [21, 23]. This is especially a problem today in the Scottish highlands where grouse estates form an integral part of the local economy. In mainland Britain, LI virus occurs speci?cally in areas of upland sheep grazing, which include Scotland and the North Yorkshire moors and is transmitted by the 3-host ixodid tick species Ixodes ricinus [22]. Until recently sheep were the only species regarded as essential for the transmission and persistence of LI virus. However, recent studies have demonstrated the ef?cient transm