Real-time Quantitative RT-PCR Assay for Quick Detection of SARS-associated Coronavirus RNA

FENG Yan*; LU Yi-yu; YAN Ju-ying; SHI Wen; LI Min-hong; GONG Li-ming; GE Qiong; ZHOU Min

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June 2005

FENG Yan*, LU Yi-yu, YAN Ju-ying, SHI Wen, LI Min-hong, GONG Li-ming, GE Qiong and ZHOU Min. Real-time Quantitative RT-PCR Assay for Quick Detection of SARS-associated Coronavirus RNA[J]. Virologica Sinica, 2005, 20(3): 228-231.

Citation: FENG Yan*, LU Yi-yu, YAN Ju-ying, SHI Wen, LI Min-hong, GONG Li-ming, GE Qiong, ZHOU Min. Real-time Quantitative RT-PCR Assay for Quick Detection of SARS-associated Coronavirus RNA .VIROLOGICA SINICA, 2005, 20(3) : 228-231.

荧光定量RT-PCR技术快速检测SARS病毒核酸

1.
浙江省疾病预防控制中心浙江杭州

摘要

建立以特异性荧光探针为特点的TaqMan荧光定量RTPCR方法用于检测严重急性呼吸道综合症病毒(severeacuterespiratorysyndromeassociatecoronavirus,SARSCoV)核酸。筛选针对SARS病毒基因保守区域设计的引物与TaqMan探针,并对荧光定量RTPCR反应体系与反应条件进行优化,验证本方法的特异性、敏感度与重复性。实验结果表明:本方法对SARS病毒核酸的检测具有高度特异性,与甲1型、甲3型、乙型流感病毒、禽流感病毒H5N1、麻疹及其他呼吸道病毒均无交叉反应;检测灵敏度达0.1TCID50;从病毒核酸提取至检测完成仅需3h左右,且操作简便,重复性好。本研究建立的TaqMan荧光定量RTPCR方法特异、敏感、快速,适合于临床实验室进行SARS病毒的早期快速检测。
- 荧光定量RTPCR
- , TaqMan探针
- , SARS病毒

Real-time Quantitative RT-PCR Assay for Quick Detection of SARS-associated Coronavirus RNA

Abstract

A real-time TaqMan-based RT-PCR assay was developed to rapidly detect the Severe acute respiratory syndrome-associate coronavirus(SARS-CoV). Primers and probes specific to the conserved region of the SARS-CoV genome were selected,and the reactive system and conditions were optimized to improve the sensitivity、specificity and repetition of the assay.The results showed that the real-time RT-PCR assay was specific and there were no cross reactions with influenza A-1,A-3,B and H-5N-1 virus、measles virus and other common respiratory viruses.The sensitivity of the assay was 0.1TCID-{50}.It took only three hours from viral RNA extraction to complete the real-time PCR,and the assay was simple and had good repeats. This real-time RT-PCR TaqMan-based assay was a specific、sensitive and quick tool suitable for early and quick detection of SARS-CoV in clinical labs.
- Real-time quantitative RT-PCR
- , TaqMan-based probe
- , SARS-CoV

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Real-time Quantitative RT-PCR Assay for Quick Detection of SARS-associated Coronavirus RNA

Abstract: A real-time TaqMan-based RT-PCR assay was developed to rapidly detect the Severe acute respiratory syndrome-associate coronavirus(SARS-CoV). Primers and probes specific to the conserved region of the SARS-CoV genome were selected,and the reactive system and conditions were optimized to improve the sensitivity、specificity and repetition of the assay.The results showed that the real-time RT-PCR assay was specific and there were no cross reactions with influenza A-1,A-3,B and H-5N-1 virus、measles virus and other common respiratory viruses.The sensitivity of the assay was 0.1TCID-{50}.It took only three hours from viral RNA extraction to complete the real-time PCR,and the assay was simple and had good repeats. This real-time RT-PCR TaqMan-based assay was a specific、sensitive and quick tool suitable for early and quick detection of SARS-CoV in clinical labs.