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当前位置: 首页 > 文章 > 马泰勒虫二温式PCR检测方法的建立及应用 中国动物检疫 2014,31 (5) 78-80+86
Position: Home > Articles > Establishment and application of a two-temperature PCR assay for detection of Theileria equi China Animal Health Inspection 2014,31 (5) 78-80+86

马泰勒虫二温式PCR检测方法的建立及应用

作  者：
李佳;李永畅;王振宝;张杨;巴音查汗
单  位：
伊犁出入境检验检疫局;新疆农业大学动物医学学院
关键词：
马泰勒虫;18S rRNA;二温式PCR;检测
摘  要：
为给马泰勒虫提供特异、快速、敏感的检测方法,根据马泰勒虫18S rRNA保守基因序列,设计了一对特异性引物,建立马泰勒虫二温式PCR检测方法。扩增出的目的片段大小为139 bp,经克隆、测序分析,与GenBank上登录的序列相似性为100%。该方法与其他虫种无交叉反应,具有一定的灵敏性和特异性。应用所建立的方法对采集的30份疑似病例全血样品进行检测,阳性率为33.3%,相比之下相同样品的血液涂片检查的阳性率为13.3%。与三温PCR方法比较减少了反应时间的消耗,提高了检测效率,该方法对马泰勒虫病的早期(潜伏感染期)检测、诊断具有实际应用价值。
译  名：
Establishment and application of a two-temperature PCR assay for detection of Theileria equi
作  者：
Li Jia;Li Yongchang;Wang Zhenbao;Zhang Yang;Bayin Chahan;College of Animal Medicine,Xinjiang Agriculture University;Yili Entry-Exit Inspection and Quarantine Bureau;
关键词：
T.equi;;18S rRNA;;two-temperature PCR;;detec
摘  要：
To establish a specifi c, rapid and sensitive detection method for Theileria equi(T.equi), a pair of specific primers was designed according to the conservative 18S rRNA gene sequence of T.equi and a two-temperature PCR assay for T.equi was established. The amplifi ed target fragment,139 bp in size was cloned and sequenced. Compared with the GenBank registered sequence,the similarity was 100% by sequence analysis.The assay showed no cross-reaction with other protozoon parasites,suggesting high specifi city. 30 whole blood samples collected were tested by the developed assay,and the positive rate of T.equi was 33.3%.While the positive rate of blood smear test was 13.3%.Compared with the three-temperature PCR assay, the reaction time was reduced and the detection effi ciency improved.The twotemperature PCR assay could be used for early detection and diagnosis of latent infection by T.equi with practical value.

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