基孔肯雅病毒荧光定量PCR检测方法的建立

中国媒介生物学及控制杂志 ›› 2012, Vol. 23 ›› Issue (1): 25-28.

基孔肯雅病毒荧光定量PCR检测方法的建立

杨宇, 白琳, 胡健萍, 姚李四, 魏莲, 杨志红, 王静

中国检验检疫科学研究院卫生检疫研究所,北京 100123

收稿日期:2011-09-09 出版日期:2012-02-20 发布日期:2012-02-20
通讯作者: 王静,Email: wangjing0115@126.com
作者简介:杨宇(1976-),女,博士,副研究员,从事病原生物学研究。Email: redyy99@gmail.com
基金资助:
国家自然科学基金(30900053);中国检验检疫科学研究院科研资金项目(2009JK027);财政部质检公益项目(2007GYJ023,2007GYJ024)

Real-time PCR method for rapid detection of Chikungunya virus

YANG Yu, BAI Lin, HU Jian-ping, YAO Li-si, WEI Lian, YANG Zhi-hong, WANG Jing

Institute of Health Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100123, China

Received:2011-09-09 Online:2012-02-20 Published:2012-02-20
Supported by:
Supported by the National Natural Science Foundation of China(No. 30900053), CIAQ Research Grant(No. 2009JK027)and Chinese Public Walfare Program(No. 2007GYJ023, 2007GYJ024)

摘要/Abstract

摘要：

目的建立一种快速、敏感、特异的实时荧光定量PCR方法,检测基孔肯雅病毒。方法通过序列比对挑选出基孔肯雅病毒基因组中高度保守的序列,在此序列上设计引物及TaqMan探针,建立实时荧光定量PCR反应体系。结果经优化的荧光定量PCR方法有较好的灵敏度和特异性,对阳性对照质粒标准品的灵敏度可达21拷贝/μl,通过检测与传播媒介相似的流行性乙型脑炎病毒、黄热病毒、登革热病毒无交叉反应。结论该方法的建立在基孔肯雅热的疾病防控方面有较好的应用前景。

关键词: 荧光定量PCR, 基孔肯雅病毒, 检测, 交叉反应

Abstract:

Objective To establish a rapid, sensitive and specific detection method for detecting Chikungunya virus(CHIKV) using Real-time fluorescence quantitative PCR. Methods With specifically designed primers and a TaqMan probe on the highly conserved sequence of CHIKV through alignment, the sensitivity of the Real-time fluorescence quantitative PCR assay was optimized by evaluating different concentrations of primers and probes. Results A specific Real - time PCR method was developed with the sensitivity of 21 copies/μl for CHIKV, a synthetic CHIKV genome DNA as a positive control; Japanese encephalitis virus, Yellow fever virus, Dengue virus were using to examine the specificity. Conclusion Promising prospects of this assay could be expected for Chikungunya fever prevention and control.

Key words: Real-time PCR, Chikungunya virus, Detection, Cross reaction

中图分类号:

R373.3

杨宇, 白琳, 胡健萍, 姚李四, 魏莲, 杨志红, 王静. 基孔肯雅病毒荧光定量PCR检测方法的建立[J]. 中国媒介生物学及控制杂志, 2012, 23(1): 25-28.

YANG Yu, BAI Lin, HU Jian-ping, YAO Li-si, WEI Lian, YANG Zhi-hong, WANG Jing. Real-time PCR method for rapid detection of Chikungunya virus[J]. Chines Journal of Vector Biology and Control, 2012, 23(1): 25-28.

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