牛副流感3型病毒Nano-PCR、LAMP方法的建立及初步应用

doi:10.16431/j.cnki.1671-7236.2017.10.002

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (10): 2837-2844.doi: 10.16431/j.cnki.1671-7236.2017.10.002

牛副流感3型病毒Nano-PCR、LAMP方法的建立及初步应用

李健友^1,2, 李家伟³, 王超², 郭利², 何洪彬¹

1. 东北农业大学, 哈尔滨 150030;
2. 中国农业科学院特产研究所, 农业部经济动物疫病重点实验室, 长春 130122;
3. 安图县畜牧兽医总站, 安图 133600

收稿日期:2017-03-07 出版日期:2017-10-20 发布日期:2017-10-20
通讯作者: 郭利, 何洪彬 E-mail:piaogl110@163.com;hongbinhe@sdnu.edu.cn
作者简介:李健友(1991-),男,山东临沂人,硕士生,研究方向:分子病毒学,E-mail:cctcs_ljy@163.com
基金资助:
国家重点研发计划：畜禽重大疫病防控于高效安全养殖综合技术研发（2016YFD0500907）；吉林省科技厅重点科技成果转化项目（20150307024NY）；吉林省科技厅产业技术创新战略联盟项目（20170309002NY）

Establishment and Preliminary Application of Nano-PCR and LAMP Methods for Bovine Parainfluenza Type-3 Virus

LI Jian-you^1,2, LI Jia-wei³, WANG Chao², GUO Li², HE Hong-bin¹

1. Northeast Agricultural University, Harbin 150030, China;
2. Key Laboratory of Special Animal Epidemic Disease, Ministry of Agricultural, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China;
3. Antu Animal Husbandry and Veterinary Station, Antu 133600, China

Received:2017-03-07 Online:2017-10-20 Published:2017-10-20

摘要/Abstract

摘要：

试验旨在建立牛副流感3型病毒（bovine parainfluenza type-3 virus，BPIV3）纳米PCR（Nano-PCR）与环介导等温扩增技术（loop-mediated isothermal amplification，LAMP）新型快速分子检测技术，对其进行特异性和敏感性对比试验，并对10份临床样品进行了检测。特异性与敏感性试验结果显示，建立的Nano-PCR和LAMP方法只对BPIV3特异，而对牛呼吸道合胞体病毒、牛传染性鼻气管炎病毒、牛病毒性腹泻病毒无交叉反应；建立的Nano-PCR和LAMP方法具有相同的敏感性，均是普通PCR的10倍，最低核酸检出量均为4.16×10²拷贝/μL。临床检测结果显示，建立的两种方法阳性符合率为100%，且阳性检出率均高于普通PCR。因此，本试验建立的Nano-PCR和LAMP方法为BPIV3的临床诊断提供了更快速、敏感、可靠的工具。

关键词: 牛副流感3型病毒(BPIV3); Nano-PCR; LAMP

Abstract:

This study was aimed to establish Nano-PCR and LAMP which were new rapid type of molecular detection technologies of bovine parainfluenza type-3 virus (BPIV3).The comparative specificity and sensitivity of BPIV3 Nano-PCR and LAMP PCR were tested, and the assay was applied to detect 10 clinical samples. The specificity and sensitivity tests showed that Nano-PCR and LAMP were only sensitive to BPIV3,without cross reaction to other viruses, such as bovine respiratory syncytial virus,infectious bovine rhinotracheitis virus and bovine viral diarrhea virus. In the sensitivity test, Nano-PCR and LAMP showed 10 times sensitivity than that of the traditional PCR technology,with the minimum detection of 4.16×10² copies/μL. Clinical test results showed that the coincidence rate of Nano-PCR and LAMP could reach to 100%, and the positive detection rate was much higher than that of normal PCR.Therefore, the Nano-PCR and LAMP methods established in this study provided a faster, sensitive and reliable tool for the clinical diagnosis of BPIV3.

Key words: bovine parainfluenza type-3 virus (BPIV3); Nano-PCR; LAMP

中图分类号:

S858.23

李健友, 李家伟, 王超, 郭利, 何洪彬. 牛副流感3型病毒Nano-PCR、LAMP方法的建立及初步应用[J]. 《中国畜牧兽医》, 2017, 44(10): 2837-2844.

LI Jian-you, LI Jia-wei, WANG Chao, GUO Li, HE Hong-bin. Establishment and Preliminary Application of Nano-PCR and LAMP Methods for Bovine Parainfluenza Type-3 Virus[J]. , 2017, 44(10): 2837-2844.

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牛副流感3型病毒Nano-PCR、LAMP方法的建立及初步应用

Establishment and Preliminary Application of Nano-PCR and LAMP Methods for Bovine Parainfluenza Type-3 Virus

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