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当前位置: 首页 > 文章 > 荧光定量PCR检测硬蜱体内莱姆病螺旋体的研究 畜牧与兽医 2010,42 (1) 23-25
Position: Home > Articles > Detection of Borrelia burgdorferi in tick by a real-time fluorescence quantitative PCR Animal Husbandry & Veterinary Medicine 2010,42 (1) 23-25

荧光定量PCR检测硬蜱体内莱姆病螺旋体的研究

作  者：
曹杰;仝彩玲;周勇志;张厚双;周金林
单  位：
中国农业科学院上海兽医研究所农业部动物寄生虫学重点实验室
关键词：
硬蜱;莱姆病;伯氏疏螺旋体;荧光定量PCR
摘  要：
为建立一个荧光定量PCR检测硬蜱体内莱姆病螺旋体的方法,根据GenBank登录的莱姆病螺旋体鞭毛蛋白FlaB序列,应用生物学软件进行序列比对,在保守的C段区设计与筛选特异引物和TaqMan探针。对荧光定量PCR反应体系与条件进行优化,验证方法的特异性、敏感性,并通过对感染螺旋体的蜱样本的检测,评价该方法的实用价值。结果显示,自然感染莱姆病螺旋体的35份蜱标本检测阳性符合率100%,正常蜱20份标本的检测结果均为阴性。该方法对牛巴贝斯原虫、泰勒原虫、边缘无浆体、金龟子绿僵菌、大肠杆菌等蜱体常见病原微生物所抽提的DNA的检测均呈阴性。荧光定量PCR方法检测质粒的灵敏度可达1×102拷贝/μL。TaqMan荧光定量PCR方法检测硬蜱体内莱姆病螺旋体具有较好的敏感性和特异性,可适于莱姆病的流行病学调查和监控。
译  名：
Detection of Borrelia burgdorferi in tick by a real-time fluorescence quantitative PCR
作  者：
CAO Jie, TONG Cai-ling, ZHOU Yong-zhi, ZHANG Hou-shuang, ZHOU Jin-lin (Shanghai Veterinary Research Institute, CAAS, Key Laboratory of Animal Parasitology of Ministry of Agriculture of China, Shanghai 200241, China)
关键词：
tick; Lyme disease; Borrelia burgdorferi; real-time PCR
摘  要：
To establish a real-time fluorescence quantitative PCR assay for detecting Borrelia burgdorferi in tick, according to the published flagellin gene sequences of B.burgdorferi, a multiple sequence alignment was done to search for primers and the fluorogenic TaqMan probe. The sensitivity and specificity of the real-time PCR assay were confirmed using the infected tick samples. The results showed that all the 35 samples from the infected ticks were positive, whereas 20 normal ticks were all negative. The samples with other pathogens, such as Babesia bovis, Theileria annulata, Anaplasma marginale, Metarhizium anisopliae and Escherichia coli, were negative. The sensitivity of the real-time PCR assay was 1×102 copies/μL of recombinant plasmid. Taken together, we have established a real-time PCR assay for detection of pathogen for Lyme disease in tick. This assay will be useful in the investigation and control of Lyme disease.

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