An internally controlled, one-step, real-time RT-PCR assay for norovirus detection and genogrouping

详细信息	查看全文 | 推荐本文 |

• 作者：K.J. Rolfe ; S. Parmar ; D. Mururi ; T.G. Wreghitt ; H. Jalal ; H. Zhang ; M.D. Curran
• 关键词：Norovirus ; Internal control ; Real-time RT-PCR ; MS2 bacteriophage ; Rotor-Gene ; Genogrouping
• 刊名：Journal of Clinical Virology
• 出版年：2007
• 期刊代码：150_13866532
• 类别：med
• 出版时间：August 2007
• 卷：39
• 期：4
• 页码：318-321
• 文件大小：376 K

摘要

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

<p>To further develop a one-step RT-PCR assay for norovirus detection/genogrouping by addition of MS2 bacteriophage as an internal control.

Study design

<p>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 MS2 phage and MS2 primers/probe on the sensitivity/specificity of the PCR assay was examined.

Results

<p>The addition of MS2 as an internal control showed no loss of sensitivity or specificity for norovirus detection.

Conclusions

<p>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.