Evaluation of a new real-time PCR assay for detection of Borrelia burgdorferi in rodents
GENG Zhen, HOU Xue-xia, ZHANG Lin, HAO Qin
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
Abstract:We established and evaluated a real-time PCR assay for detection of Borrelia burgdorferi (B. burgdorferi) in rodents. MGB probe and specific primers were designed based on the conserved sequences of recA gene of B. burgdorferi and methodology evaluation was performed. The 123 rodent tissue samples were detected by the established real-time PCR and nested PCR methods. Results showed real-time PCR assay based on recA gene was successfully established. Specificity evaluation showed specific amplification was only achieved from genomic DNA of B. burgdorferi. The lowest detection limit of the new assay was about 10 copies of recA gene from B. burgdorferi genomic DNA. The average intra-and-inter coefficient of variations (CV) of the Ct value were 1.56% and 2.30% respectively. In 123 rodent samples, 59 samples were detected positive by real-time PCR, compared to 43 positives by nested PCR. The new real-time PCR assay is suitable for detecting Borrelia burgdorferi in rodents.
耿震, 侯学霞, 张琳, 郝琴. 一种莱姆病螺旋体real-time PCR方法的建立及其在鼠标本检测中的应用评价[J]. 中国人兽共患病学报, 2015, 31(9): 812-816.
GENG Zhen, HOU Xue-xia, ZHANG Lin, HAO Qin. Evaluation of a new real-time PCR assay for detection of Borrelia burgdorferi in rodents. Chinese Journal of Zoonoses, 2015, 31(9): 812-816.
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