Development of a nested PCR assay to detect equine infectious anemia proviral DNA from peripheral blood of naturally infected horses.
Equine infectious anemia (EIA) has posed a major challenge and caused significant losses to the equine industry worldwide. PCR detection methods have considerable potential as an adjunct to conventional serological diagnostic techniques. However, most published PCR methods, including that recommended by the OIE, were designed using laboratory-adapted virus strains and do not function with field isolates of EIA virus (EIAV). In the present study, a nested PCR assay for detection of EIAV proviral DNA in peripheral blood cells of naturally infected horses was developed. Primer sets were designed based on conserved 5' regions of the viral genome extending from the LTR to the tat gene. Preliminary studies demonstrated that the method has a detection limit of 10 genomic copies and, when applied to a naturally EIAV-infected feral horse population, shows 100 % correlation with conventional serological diagnostic techniques. This assay provides a powerful new tool in the control of EIAV.
Department of Veterinary Microbiology, University of Miyazaki, 1-1 Gakuen Kibanadai Nishi, Miyazaki 889-2192, Japan.
Jian-Bao
JB
Dong
Wei
W
Zhu
Frank R
FR
Cook
Yoshitaka
Y
Goto
Yoichiro
Y
Horii
Takeshi
T
Haga
D000818:Animals; D001769:Blood / Q000821:virology*; D017124:Conserved Sequence; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004859:Equine Infectious Anemia / Q000175:diagnosis* / Q000821:virology; D006736:Horses; D004860:Infectious Anemia Virus, Equine / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014730:Veterinary Medicine / Q000379:methods*
D017931:DNA Primers; D004279:DNA, Viral