title: Development of real-time polymerase chain reaction for detection of Borrelia burgdorferi sensu lato in China.
abstract: Universal primers and probes were selected on the basis of the 16S rRNA gene sequence of Borrelia burgdorferi in GenBank®, and a real-time polymerase chain reaction (PCR) method for detection of B. burgdorferi was established. The results showed that this method could specifically detect the B31 strain (Borrelia burgdorferi sensu stricto), the BO23 strain (Borrelia afzelii), and the SZ strain (Borrelia garinii), without cross-reaction with genome DNA of Theileria (T. luwenshuni, T. uilenbergi, T. sinensis, T. annulata, T. sergenti, T. annulata), Babesia (B. bigemina, B. ovate, B. sp. (Xinjiang)), Anaplasma (A. marginale, A. ovis), Mycoplasma mycoides subsp. capri, and Chlamydia psittaci, which are the infective pathogens to yak and/or sheep. The sensitivity of this real-time PCR is 10⁴ times greater than that of a conventional PCR. The real-time PCR was able to amplify 16S rRNA gene from as few as 22.88 fg genomic DNA of B. burgdorferi sensu lato. Tick DNAs from 369 field samples collected from Shangzhi City of Heilongjiang Province were tested, resulting in an infection rate of 42.80%, and a total of 332 genomic DNAs from the blood of 186 yaks and 146 sheep in the Gannan Tibetan Autonomous Prefecture of Gansu Province were tested, resulting in 24.19% positive rate for the yaks and 39.04% positive rate for the sheep.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases MOA, Key Laboratory of Veterinary Parasitology of Gansu Province, Chinese Academy of Agricultural Sciences, Lanzhou Veterinary Research Institute, Lanzhou, P.R. China.
authors.forename: Yuguang
authors.initials: Y
authors.lastname: Fu
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Liu
authors.forename: Guiquan
authors.initials: G
authors.lastname: Guan
authors.forename: Qingli
authors.initials: Q
authors.lastname: Niu
authors.forename: Youquan
authors.initials: Y
authors.lastname: Li
authors.forename: Jifei
authors.initials: J
authors.lastname: Yang
authors.forename: Qiaoyun
authors.initials: Q
authors.lastname: Ren
authors.forename: Miling
authors.initials: M
authors.lastname: Ma
authors.forename: Aihong
authors.initials: A
authors.lastname: Liu
authors.forename: Yulv
authors.initials: Y
authors.lastname: Peng
authors.forename: Jianxun
authors.initials: J
authors.lastname: Luo
authors.forename: Hong
authors.initials: H
authors.lastname: Yin
mesh_terms: D000818:Animals; D015748:Borrelia burgdorferi Group / Q000235:genetics* / Q000302:isolation & purification*; D002417:Cattle; D002418:Cattle Diseases / Q000453:epidemiology / Q000382:microbiology; D002681:China / Q000453:epidemiology; D004269:DNA, Bacterial / Q000235:genetics*; D003870:Dermacentor / Q000382:microbiology; D016680:Genome, Bacterial; D008193:Lyme Disease / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000453:epidemiology / Q000382:microbiology
chemical_list: D004269:DNA, Bacterial