Development of a TaqMan Real-time PCR assay for detection of Batai virus

doi:10.11853/j.issn.1003.4692.2016.01.001

Chines Journal of Vector Biology and Control ›› 2016, Vol. 27 ›› Issue (1): 1-4.DOI: 10.11853/j.issn.1003.4692.2016.01.001

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Development of a TaqMan Real-time PCR assay for detection of Batai virus

CAO Yu-xi, HE Xiao-xia, FU Shi-hong, LI Hao, LIANG Guo-dong, WANG Huan-yu

National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China

Received:2015-11-23 Online:2016-02-20 Published:2016-02-20
Supported by:
Supported by the National Science and Technology Major Project of the Ministry of Science and Technology of China (No. 2013ZX10004-101)

巴泰病毒实时荧光定量PCR检测方法的建立

曹玉玺, 赫晓霞, 付士红, 李浩, 梁国栋, 王环宇

中国疾病预防控制中心病毒病预防控制所, 北京 102206

通讯作者: 王环宇,Email:rainoffall@yahoo.com
作者简介:曹玉玺,男,博士,副主任技师,主要从事虫媒病毒病防控和实验室生物安全管理工作,Email:yuxicao@hotmail.com
基金资助:
国家科技重大专项课题(2013ZX10004-101)

Abstract

Abstract:

Objective To develop a rapid and sensitive detection method for Batai virus (BATV) based on TaqMan Real-time PCR. Methods Based on the BATV NS gene sequences of S segment published in GenBank, BATV specific primers and probe were designed. The specificity and stability of the system were evaluated. Quantitative standard curve of BATV TaqMan Real-time PCR was established. Results The specificity and stability test showed that the system is specific and the coefficient variables were all less than 2.50%. Quantitative standard curve based on the genomic copy was drawn, and the lowest detectable limit (LOD) of system is 10 copies/μl. Conclusion TaqMan Real-time PCR for BATV detection has been developed, which is more sensitive and more efficient than the general PCR.

Key words: Batai virus, TaqMan Real-time PCR, Molecular detection

摘要：

目的利用TaqMan 实时荧光定量PCR技术,建立巴泰病毒(Batai virus,BATV)实时荧光定量PCR检测方法。方法根据GenBank发表的BATV基因序列资料分析结果,在其S片段编码非结构蛋白的基因区段设计BATV特异引物与探针,利用不同科、属的13株病毒核酸验证方法特异性,利用体外转录的定量RNA标准品建立基因拷贝数定量分析模型,重复实验检验方法的稳定性。结果引物与探针具有良好的特异性,定量分析模型的灵敏度为10拷贝/μl,同一样品重复检测循环阈值的变异系数均<2.50%。结论建立了一种特异、灵敏、高效的BATV一步法TaqMan 实时荧光定量PCR检测方法,为今后该病毒的检测与研究工作提供技术手段。

关键词: 巴泰病毒, 实时荧光定量PCR, 分子检测

CLC Number:

R373.3

CAO Yu-xi, HE Xiao-xia, FU Shi-hong, LI Hao, LIANG Guo-dong, WANG Huan-yu. Development of a TaqMan Real-time PCR assay for detection of Batai virus[J]. Chines Journal of Vector Biology and Control, 2016, 27(1): 1-4.

曹玉玺, 赫晓霞, 付士红, 李浩, 梁国栋, 王环宇. 巴泰病毒实时荧光定量PCR检测方法的建立[J]. 中国媒介生物学及控制杂志, 2016, 27(1): 1-4.

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Development of a TaqMan Real-time PCR assay for detection of Batai virus

巴泰病毒实时荧光定量PCR检测方法的建立

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