An internally controlled, one-step, real-time RT-PCR assay for norovirus detection and genogrouping
An internally controlled, one-step, real-time RT-PCR assay for norovirus detection and genogrouping
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DOI：
10.1016/j.jcv.2007.05.005
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发表时间：
2007-08-01
影响因子：
8.8
通讯作者：
Curran, M. D.
中科院分区：
文献类型：
--
作者：
Rolfe, K. J.;Parmar, S.;Curran, M. D.
背景：由于粪便中的抑制物质，用于诺如病毒检测的逆转录（RT）-PCR容易出现假阴性结果。需要一个内部控制，以监测提取效率和检测inhibit 1 n.Objectives：To further developed a one-step RT-PCR assay for norovirus detection/genogrouping by addition of MS 2 bacteriophage as an internal control.Studydesign：我们的诺如病毒RT-PCR assay进行了修改，通过添加MS 2噬菌体的提取托盘和引物/探针MS 2检测的反应混合物。结果：添加MS 2作为内对照，诺如病毒检测的灵敏度和特异性均没有降低。结论：建立了一种以MS 2为内对照的三重、一步法、特异性、实时荧光定量RT-PCR方法，可用于诺如病毒感染的常规实验室诊断。(c)2007 Elsevier BX版权所有。
Background: Reverse transcription (RT)-PCR for norovirus detection is prone to false-negative results due to inhibitory substances in faeces. An internal control is needed to monitor extraction efficiency and to detect inhibition.Objectives: To further develop a one-step RT-PCR assay for norovirus detection/genogrouping by addition of MS2 bacteriophage as an internal control.Studydesign: Our norovirus RT-PCR assay was modified by addition of MS2 phage to the extraction tray and primers/probe for MS2 detection to the reaction mix. The effect of addition of NIS2 phage and MS2 primers/probe on the sensitivity/specificity of the PCR assay was examined.Results: The addition of MS2 as an internal control showed no loss of sensitivity or specificity for norovirus detection.Conclusions: A triplex, one-step, type-specific, real-time RT-PCR with MS2 internal control has been developed for use in routine laboratory diagnosis of norovirus infection. (c) 2007 Elsevier BX All rights reserved.