动物衣原体RPA检测方法的建立  

Detection of Chlamydia in livestock by recombine polymerase amplification

作　　者：费媛媛[1] 李兆才[1] 娄忠子[1] Muhammed MUSTAFA 张志君 赵宏涛[2] 祁世荣 李长云[4] 周继章[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃兰州730046 [2]陕西省动物疫病预防控制中心,陕西西安712100 [3]青海省都兰县宗家乡兽医站,青海都兰816100 [4]青海省海晏县畜牧兽医站,青海海晏812200

出　　处：《中国兽医科学》2018年第2期142-147,共6页Chinese Veterinary Science

基　　金：“十三五”国家重点研发计划“牛羊重要病原体分子检测新技术研究”(2016YFD0500907)

摘　　要：为建立一种快速、准确的动物衣原体检测方法,本研究基于动物衣原体的保守基因env B的序列,设计并合成引物与探针,建立了一种动物衣原体特异性重组酶聚合酶扩增（RPA）检测方法。该检测方法在37-39℃恒温反应20 min,即可实现对动物衣原体目的基因片段的有效扩增,并具有高灵敏度的特点,最低检测限为101copies/L,同时利用建立的RPA检测方法和普通PCR方法 对10份临床样品进行衣原体检测,结果阳性检出率完全一致。表明,本研究建立的动物衣原体RPA检测方法操作简单方便、灵敏度高,短时间内能获得准确可靠的诊断结果,对动物衣原体的临床检测和疾病诊断提供了一种新的、可靠的技术支持。The aim of this study was to establish a rapid detection method for Chlamydia in abortive animals to facilitate timely treatment. A recombines polymerase amplification（RPA） method was developed using primers and a probe specific for the conserved gene of envB. The RPA reaction was successfully performed at 37--39℃ for 20 min. With recombinant plasmid DNAcarrying envBgene as template, the test results showed that the sensitivity of RPA was 101copies of plasmid DNA. Positive rate for Chlamydia detection of 10 clinical samples by RPA was consistent with that by conventional PCR. The established RPA method is simple,rapid,and reliable,which can be utilized as a novel tool for experimental research and detection of Chlamydial infection in clinical samples.

关 键 词：重组酶聚合酶扩增 衣原体 快速检测 

分 类 号：S852.67[农业科学—基础兽医学]