Comparison of polymerase chain reaction methods for the detection of Theileria equi infection using whole blood compared with pre-extracted DNA samples as PCR templates.
Rapid, efficient, and reproducible procedures for isolating DNA before PCR gene amplification are essential for the diagnosis of piroplasms. In this study, we evaluated the ease and reliability of detecting Theileria equi by PCR using pre-extracted DNA samples (by QIAamp DNA Mini Kit and phenol-chloroform methods) compared with blood spotted on FTA cards as PCR templates. Although minimal variations in limit of detection were observed among the methods compared, overall, the use of pre-extracted DNA samples and blood spotted on FTA cards had comparable detection limits. These results indicate that T. equi infection can be efficiently detected directly from FTA cards by PCR without the need for pre-extraction of DNA from blood samples.
Alhassan
A
A
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
Iseki
H
H
Kim
C
C
Yokoyama
N
N
Igarashi
I
I
D000818:Animals; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D005869:Ghana; D006734:Horse Diseases / Q000097:blood / Q000175:diagnosis / Q000469:parasitology*; D006736:Horses; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000737:chemistry / Q000235:genetics; D016794:Theileria / Q000235:genetics / Q000302:isolation & purification*; D013801:Theileriasis / Q000097:blood* / Q000469:parasitology
D016054:DNA, Protozoan; D015800:Protozoan Proteins; D012337:RNA, Ribosomal, 18S