State Hygienic Laboratory at the University of Iowa, Research Park, Iowa City, IA 52242, United States.
J Virol Methods. 2011 Jul;175(1):80-4. doi: 10.1016/j.jviromet.2011.04.022. Epub 2011 Apr 28.
Although norovirus has been identified as the most common cause of gastroenteritis, the majority of cases have no etiologic agent identified. In this study, we describe the optimization of a real-time RT-PCR assay for the improved detection of genogroup I norovirus in patient specimens based upon sequence data from a collection of representative clinical norovirus sequences. The redesigned assay demonstrated a 64 fold increase in sensitivity, a 2 log decrease in the limit of detection, and an 18% increase in amplification efficiency, when compared to the standard assay. The optimized test also detected GI norovirus in clinical specimens that were initially negative by the standard assay. Use of the optimized assay increased the annual positivity of GI norovirus in Iowa from 1.2% to 4.5%, indicating the prevalence of GI norovirus may be higher than previously identified. Laboratory confirmation of the etiologic agent involved in gasteroenteritis cases is essential for better understanding of the prevalence and transmission of noroviruses.
虽然诺如病毒已被确定为导致肠胃炎的最常见原因，但大多数病例的病因仍未确定。在这项研究中，我们描述了一种实时 RT-PCR 检测方法的优化，该方法基于代表性临床诺如病毒序列集的序列数据，可提高对 I 型诺如病毒属的检测。与标准检测相比，重新设计的检测方法的灵敏度提高了 64 倍，检测限降低了 2 个对数，扩增效率提高了 18%。优化后的检测方法还能检测到最初用标准检测方法呈阴性的临床标本中的 GI 诺如病毒。优化后的检测方法的使用使爱荷华州 GI 诺如病毒的年阳性率从 1.2%增加到 4.5%，表明 GI 诺如病毒的流行率可能高于之前的报告。对肠胃炎病例中涉及的病原体进行实验室确认对于更好地了解诺如病毒的流行和传播至关重要。
