Diagnostic real-time PCR assay for the quantitative detection of Theileria equi from equine blood samples.
We developed a TaqMan real-time polymerase chain reaction (PCR) assay for the quantitative detection of Theileria equi from the in vitro-cultured parasite and field blood samples collected from horses living in Ghana and Brazil. The detection limit for the assay was determined to be 1.5 parasites/microl per sample, and the quantitative capacity was demonstrated using the in vitro-cultured parasite. For field applications, the real-time PCR assay was compared to a previously established nested PCR assay used as the gold standard for the real-time PCR assay. Of 65 field blood samples, 46 samples were T. equi-positive in the nested PCR assay, while the real-time PCR assay also detected the parasite in all 46 of the nested PCR-positive samples but did not detect T. equi in the remaining 19 negative blood samples. This quantitative real-time PCR assay provides a valuable tool for fast laboratory diagnostic assessment of T. equi infection in horses.
Kim
Chul-min
CM
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
Blanco
Lidia Beatriz Conza
LB
Alhassan
Andy
A
Iseki
Hiroshi
H
Yokoyama
Naoaki
N
Xuan
Xuenan
X
Igarashi
Ikuo
I
D000818:Animals; D001938:Brazil; D017931:DNA Primers / Q000737:chemistry; D005869:Ghana; D006734:Horse Diseases / Q000175:diagnosis* / Q000469:parasitology; D006736:Horses; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity; D016794:Theileria / Q000235:genetics / Q000302:isolation & purification*; D013801:Theileriasis / Q000097:blood / Q000175:diagnosis*
D017931:DNA Primers