Rapid detection of norovirus genogroup II in clinical and environmental samples using recombinase polymerase amplification
Rapid detection of norovirus genogroup II in clinical and environmental samples using recombinase polymerase amplification
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使用重组酶聚合酶扩增快速检测临床和环境样本中的诺如病毒基因组 II
DOI：
10.1016/j.ab.2020.113834
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发表时间：
2020-09-15
影响因子：
2.9
通讯作者：
Xu, Xiangdong
中科院分区：
文献类型：
--
作者：
Han, Yanzhen;Wang, Jianchang;Xu, Xiangdong
诺如病毒是引起全球急性胃肠炎的主要病原，其流行的主要基因型是诺如病毒基因组II（NoVs GII）。NoV的快速检测很重要，因为它可以促进及时诊断。在本研究中，我们基于基因组的ORF 1-ORF 2连接处的保守区域设计了通用特异性引物和Exo探针，以与NoVs Gil的所有遗传簇杂交。本研究首次建立了一种快速、可靠的逆转录重组酶聚合酶扩增（RT-RPA）方法，可在20 min内检测出Gil型NoVs，该方法可特异性扩增出Gil型NoVs，检测限低至1.66 × 10（2）copies/mu L。在LOD、准确度和专属性方面对该方法进行了验证。我们测试了55个真实的样本，包括食物、水和粪便。结果显示对NoVs Gil的敏感性为96%，特异性为100%。整个过程可以通过移动的手提箱实验室操作，这对于资源有限的诊断实验室是有用的。这种新型的实时RT-RPA检测是一种用于NoV即时检测的准确工具，为诺如病毒引起的疾病诊断和预防提供了实际支持。
Norovirus is the leading cause of acute gastroenteritis all over the world, and the most genotype that causes its epidemic is norovirus genogroup II (NoVs GII). Rapid detection of NoVs is important because it can facilitate timely diagnosis. In this study, we designed universal specific primers and an Exo probe to hybridize to all genetic clusters of NoVs Gil based on the conserved region at the ORF1-ORF2 junction of the genome. For the first time, we established a rapid and reliable reverse transcription recombinase polymerase amplification (RT-RPA) method for the detection of NoVs Gil within 20 min. This method can specifically amplify NoVs Gil, and the detection limit was as low as 1.66 x 10(2) copies/mu L. The method was validated in terms of LOD, accuracy, and specificity. We tested 55 real samples including foods, water, and feces. The results showed a sensitivity of 96% and specificity of 100% to NoVs Gil. The whole procedure can be operated by a mobile suitcase laboratory, which is useful for resource-limited diagnostic laboratories. This novel real-time RT-RPA assay is an accurate tool for point-of-care testing of NoVs, providing practical support for norovirus-caused disease diagnosis and prevention.