| 牛轮状病毒三种核酸检测方法的比较 |
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| 引用本文： | 范晴,谢芝勋,刘加波,庞耀珊,邓显文,谢志勤,谢丽基.牛轮状病毒三种核酸检测方法的比较[J].动物检疫,2012(2):38-41. |
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| 作者姓名： | 范晴 谢芝勋 刘加波 庞耀珊 邓显文 谢志勤 谢丽基 |
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| 作者单位： | 广西兽医研究所 广西畜禽疫苗新技术重点实验室,广西南宁530001 |
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| 基金项目： | 国家百千万人才工程人选专项资金资助（945200603）; 广西科技攻关与新产品试制项目（桂科合0992033-5）; 广西特聘专家专项经费资助项目（2011B020） |
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| 摘 要： | 利用针对牛轮状病毒（BRV）的普通PCR、实时荧光定量PCR（Real-time PCR）和环介导等温扩增技术（LAMP）三种核酸检测方法对不同质粒浓度的样品和417份临床疑似样品进行检测,比较三种核酸检测方法的检出结果。三种核酸检测方法中LAMP最敏感敏感,能检测到1拷贝/μL,Real-time PCR能检测到100拷贝/μL,普通PCR只能检测到1×104拷贝/μL。但结合临床样品检测表明：常规PCR方法敏感度低会造成一部分漏检,LAMP灵敏度高又会造成错检。综合比较三种方法后,推荐用LAMP结合real-time PCR,不仅节约成本,且结果更为准确可靠,可提高牛轮状病毒的检出率。
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| 关 键 词： | 牛轮状病毒 RT-PCR 实时荧光PCR RT-LAMP |
Comparison of three nucleic acid amplification assays for bovine rotavirus |
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| Authors: | Fan Qing Xie Zhixun Liu Jiabo Pang YaoShan Deng XianWen Xie Zhiqin Xie Liji |
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| Institution: | (Guangxi Veterinary Research Institute,Guangxi Key Laboratory of Animal Vaccines and New Technology,Nanning 530001,China) |
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| Abstract: | 10-fold dilution series samples and 417 clinical samples were used in this study to evaluate the three BRV-specific detected methods： conventional PCR,real-time PCR and LAMP. The results showed that LAMP is the most sensitive method with its detection limit of 1 copies/μL,real-time PCR is 10copies/μL,but PCR had a lower sensitivity with its detection limit of 10000 copies/μL. Clinically,a contrast analysis of these three methods has learnt： the PCR had a higher undetected rate due to its low sensitivity to clinical samples;the error rate of LAMP was high due to its high sensitivity. These results suggested that the method of LAMP combining with real-time PCR is recommended in clinical detection of BRV,which is not only cost efficient but also accurate. |
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| Keywords: | bovine rotavirus RT-PCR real-time PCR RT-LAMP |
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