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当前位置: 首页 > 文章 > 猪细环病毒PCR-DHPLC检测技术的建立及应用 畜牧兽医学报 2012,43 (9) 1422-1428
Position: Home > Articles > Development and Application of PCR-DHPLC Assays for Detection of the Swine Torque Teno Sus Virus Chinese Journal of Animal and Veterinary Sciences 2012,43 (9) 1422-1428

猪细环病毒PCR-DHPLC检测技术的建立及应用

作  者：
杨春华;周延;孙思扬;钟毅;王川庆
单  位：
江西出入境检验检疫局;河南农业大学牧医工程学院
关键词：
猪细环病毒;变性高效液相色谱;实时荧光PCR;PCR
摘  要：
为了应用PCR结合变性高效液相色谱(DHPLC)技术检测猪细环病毒1型和2型,根据猪细环病毒的序列特点设计特异性引物,PCR扩增产物经DHPLC技术进行快速检测。选择猪细小病毒、猪圆环病毒Ⅱ型和猪伪狂犬病毒进行特异性试验,无交叉反应。该检测方法具有较好的特异性和重复性;用质粒标准品进行TTSuV1和TTSuV2的PCR-DHPLC检测,灵敏度可达1.0×101拷贝·μL-1。对80份血清样本分别应用实时荧光PCR法、PCR-凝胶电泳法及本文所建立的PCR-DHPLC法与进行检测,发现TTSuV1有63份PCR-DHPLC阳性,63份实时荧光PCR阳性,54份PCR-凝胶电泳阳性;TTSuV2有69份PCR-DHPLC阳性,69份实时荧光PCR阳性,56份PCR-凝胶电泳阳性。本试验建立的PCR-DHPLC法具有特异、敏感、快速、重复性好等优点,可用于TTSuV感染的分子流行病学调查。
译  名：
Development and Application of PCR-DHPLC Assays for Detection of the Swine Torque Teno Sus Virus
作  者：
YANG Chun-hua1,2,ZHOU Yan2,3,SUN Si-yang2,ZHONG Yi2,WANG Chuan-qing1(1.College of Animal Science and Veterinary Medicine,Henan Agricultural University, Zhengzhou 450002,China;2.Jiangxi Entry-Exit Inspection & Quarantine Bureau of P.R.China, Nanchang 330002,China;3.Nanchang University,Nanchang 330002,China)
关键词：
Swine Torque teno sus virus;DHPLC;real-time PCR;PCR
摘  要：
To identify the Torque teno sus virus(TTSuV),a PCR-DHPLC assay was developed in this study.Primers specific for the partial region of the TTSuV1 and TTSuV2 were selected to conduct the PCR-DHPLC assays.The specificity test was performed with PPV,PCV-Ⅱ and PRV and no cross reaction was found.The PCR-DHPLC for TTSuV had good specificity and nice repeatability.Sensitivity analysis showed that the developed PCR-DHPLC could detect 1.0×101 copy·μL-1.The established method and the Real-time PCR were used to detect 80 serum samples,the results showed that 63 samples were TTSuV1 positive by PCR-DHPLC,and it was consistent with real-time PCR test;69 samples were TTSuV2 positive by PCR-DHPLC,and it was consistent with real-time PCR test,56 samples were positive by normal RT-PCR.The method showed nice specification,sensitivity,repeatability,and quickness,and it could be used in epidemiological investigation of TTSuV.

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