[1]陈勉乔,于浩洋.禽流感病毒H7N9型 实时荧光定量RT-PCR检测方法的研究[J].现代检验医学杂志,2016,31(03):48-50.[doi:10.3969/j.issn.1671-7414.2016.03.013]
　CHEN Mian-qiao,YU Hao-yang.Devolepment of Real-time Fluorescence Quantitative RT-PCR Detection for Avian Influenza Virus H7N9 Type[J].Journal of Modern Laboratory Medicine,2016,31(03):48-50.[doi:10.3969/j.issn.1671-7414.2016.03.013]
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禽流感病毒H7N9型 实时荧光定量RT-PCR检测方法的研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第31卷
期数:
2016年03期
页码:
48-50
栏目:
论著
出版日期:
2016-06-15

文章信息/Info

Title:
Devolepment of Real-time Fluorescence Quantitative RT-PCR Detection for Avian Influenza Virus H7N9 Type
文章编号:
1671-7414(2016)03-048-04
作者:
陈勉乔; 于浩洋
深圳市晋百慧生物有限公司,广东深圳 518055
Author(s):
CHEN Mian-qiao; YU Hao-yang
Shenzhen GeneBioHealth Co.,LTD,Guangdong Shenzhen 518055,China
关键词:
禽流感病毒H7N9型;  实时荧光定量;  RT-PCR;  检测
分类号:
R373.13; Q503
DOI:
10.3969/j.issn.1671-7414.2016.03.013
文献标志码:
A
摘要:
目的 建立一种快速准确诊断禽流感病毒H7N9型的实时荧光定量RT-PCR检测方法。方法 参照国家生物技术信息中心(NCBI)公布的H7N9基因HA序列,设计特异性引物及TaqMan探针; 构建阳性重组质粒,验证其最低检测拷贝数、重复性及特异度。结果 试验构建的阳性重组质粒,线性关系在6×102 copies/μl～6×108 copies/μl范围内检测结果良好; 用该方法测得最低拷贝数为600 copies/μl,特异度和重复性(CV<1%)良好,无交叉反应; 用该荧光定量RT-PCR检测方法检测8份阳性样品和8份阴性样品,准确率为100%(16/16)。结论 实验建立的实时荧光定量RT-PCR检测方法可用于禽流感病毒H7N9型的快速检测。
Abstract:
Objective To establish a rapid and accurate method of fluorescent quantitative PCR detection for Avian Influenza Virus H7N9 type.Methods A pair of primers and TaqMan fluorescent probes were designed specificly for H7N9 gene according to the published nucleotidesequence from National Center for Biotechnology Information.Positive recombinant plasmid was built and the minimum copies of detection,repeatability and specificitv were tested.Results The results showed that this assay obtained positive recombinant plasmid,the linear relation was fine in the range from 102 to 108 copies/μl.The minimum copies was 500 copies/μl with this method,specificitv and repeatability(CV<1%)were fine,no cross reaction with other viruses.The precision was100%(16/16)with this method for detecting 8 positive samples and 8 negativesamples.Conclution The establishment methods of fluorescent quantitative PCR detection could be used for the rapid diagnosis Avian Influenza virus H7N9 types.

参考文献/References:

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备注/Memo

备注/Memo:
基金项目:深圳市技术创新计划(CYZZ20140605154742155); 深圳市南山技术研发和创意设计项目(KC2014JSJS0018A)。 作者简介:陈勉乔(1983-),男,硕士研究生,主要从事疾病快速诊断试剂盒的开发,Tel:15914069189,E-mail:mianqiaochen@yeah.net。 通讯作者:于浩洋(1971-),男,德籍博士,高级工程师,深圳大学客座教授,主要从事医学分子学研究,E-mail:guixuanyu@hotmail.com。
更新日期/Last Update: 2016-06-25