Definitive identification of louping ill virus by RT-PCR and sequencing in field populations of Ixodes ricinus on the Lochindorb estate.
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 identification of louping ill (LI) virus in field 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 field specimens. Development of this system should improve the capacity to undertake detailed epizootiological studies of LI virus.
Gaunt
M W
MW
NERC, Institute of Virology and Environmental Microbiology, Oxford, UK.
Jones
L D
LD
Laurenson
K
K
Hudson
P J
PJ
Reid
H W
HW
Gould
E A
EA
D000818:Animals; D000956:Antigens, Viral; D001483:Base Sequence; D006224:Cricetinae; D004279:DNA, Viral; D004669:Encephalitis Viruses, Tick-Borne / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D019084:Fluorescent Antibody Technique, Indirect; D014644:Genetic Variation; D018884:Ixodes / Q000821:virology*; D008647:Mesocricetus; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014759:Viral Envelope Proteins / Q000235:genetics
D000956:Antigens, Viral; D004279:DNA, Viral; D014759:Viral Envelope Proteins; C076634:NEG envelope glycoprotein, Negishi virus