Development of a Polymerase Chain Reaction Method for Diagnosing Babesia gibsoni Infection in Dogs

Journal of Veterinary Medical Science

Ikuo IGARASHI¹  Takeshi MIKAMI¹  Kozo FUJISAKI¹  Shinya SHIGENO²  Elikira KIMBITA¹  Xuenan XUAN¹  Hideyuki NAGASAWA¹  Shinya FUKUMOTO¹ 

[1] National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine;Leo Animal Hospital

关键词: Babesia canis;    Babesia gibsoni;    IFAT;    PCR;    P18 gene;   

DOI  :  10.1292/jvms.63.977

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(20)Cited-By(35)A pair of oligonucleotide primers were designed according to the nucleotide sequence of the P18 gene of Babesia gibsoni (B. gibsoni), NRCPD strain, and were used to detect parasite DNA from blood samples of B. gibsoni-infected dogs by polymerase chain reaction (PCR). PCR was specific for B. gibsoni since no amplification was detected with DNA from B. canis or normal dog leucocytes. PCR was sensitive enough to detect parasite DNA from 2.5 μl of blood samples with a parasitemia of 0.000002%. PCR detected parasite DNA from 2 to 222 days post-infection in sequential blood samples derived from a dog experimentally infected with B. gibsoni. The detection of B. gibsoni DNA by PCR was much earlier than the detection of antibodies to B. gibsoni in blood samples by the indirect fluorescent antibody test (IFAT) or that of the parasite itself in Giemsa-stained thin blood smear film examined by microscopy. In addition, 28 field samples collected from dogs in Kansai area, Japan, were tested for B. gibsoni infection. Nine samples were positive in blood smears, 9 samples were positive by IFAT and 11 samples were positive for B. gibsoni DNA by PCR. The nucleotide sequences of PCR products from all 11 samples found positive by PCR were completely identical to that of the P18 gene of the B. gibsoni, NRCPD strain. These results suggest that PCR provides a useful diagnostic tool for the detection of B. gibsoni infection in dogs.

【 授权许可】

Unknown