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.pdfVIP

中国人兽共患病学报 812 Chinese ]ournal o f Zoonoses 2015 , 31 (9) 001: 10. 3969/j. ìssn. 1002 - 2694 .2015. 09. 006 Evaluation of a new real-time PCR assay for detection of Borrelia burgdorf eri in rodents GENG Zhen ,HOU Xue- xia ,ZHANG Lin ,HAO Qin (State Key Laboratory for Infectious Disease Prevention and ControL. NationaL lnstitute for Communicable Disease ControL and Prevention. Chinese Center for Disease ControL and Prevention. Beijing 102206. China) Abslract ,We established and evaluated a real-time PCR assay for detection of Borrelia 阳rgdorferi (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 nes- tcd PCR methods. Rcsults showed real-time PCR assay based on recA gene was succc.ssfully 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 varia- tions (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 burg- dorferi in rodents. Keywords : BorreLia burgdorferi; real-time PCR; nested PCR; rodent Supported by the 12th Fiv~Year Major National Science and Technology Projects of China (Nos. 2012ZXI0004219-007 and 2013ZXI0004001 ) Corresp