SARS-CoV-2 监测系统升级:新基因检测方法的部署。
An Upgrade on the Surveillance System of SARS-CoV-2: Deployment of New Methods for Genetic Inspection.
机构信息
Institute for Research in Biomedical Sciences (IICB), University Center for Health Sciences, University of Guadalajara, Guadalajara 44340, Mexico.
Translational Institute of Genomic Singularity (ITRASIG), Irapuato 36615, Mexico.
出版信息
Int J Mol Sci. 2022 Mar 15;23(6):3143. doi: 10.3390/ijms23063143.
SARS-CoV-2 variants surveillance is a worldwide task that has been approached with techniques such as Next Generation Sequencing (NGS); however, this technology is not widely available in developing countries because of the lack of equipment and limited funding in science. An option is to deploy a RT-qPCR screening test which aids in the analysis of a higher number of samples, in a shorter time and at a lower cost. In this study, variants present in samples positive for SARS-CoV-2 were identified with a RT-qPCR mutation screening kit and were later confirmed by NGS. A sample with an abnormal result was found with the screening test, suggesting the simultaneous presence of two viral populations with different mutations. The DRAGEN Lineage analysis identified the Delta variant, but there was no information about the other three mutations previously detected. When the sequenced data was deeply analyzed, there were reads with differential mutation patterns, that could be identified and classified in terms of relative abundance, whereas only the dominant population was reported by DRAGEN software. Since most of the software developed to analyze SARS-CoV-2 sequences was aimed at obtaining the consensus sequence quickly, the information about viral populations within a sample is scarce. Here, we present a faster and deeper SARS-CoV-2 surveillance method, from RT-qPCR screening to NGS analysis.
SARS-CoV-2 变体监测是一项全球性任务,已经采用了下一代测序(NGS)等技术来进行;然而,由于缺乏设备和科学资金有限,发展中国家无法广泛应用这项技术。一种选择是部署 RT-qPCR 筛选测试,该测试有助于在更短的时间内以更低的成本分析更多数量的样本。在这项研究中,使用 RT-qPCR 突变筛选试剂盒鉴定了 SARS-CoV-2 阳性样本中的变体,然后通过 NGS 进行了确认。筛选测试发现了一个异常结果的样本,表明同时存在两种具有不同突变的病毒群体。DRAGEN 谱系分析鉴定出了 Delta 变体,但没有关于之前检测到的其他三种突变的信息。当对测序数据进行深入分析时,发现存在具有不同突变模式的读取,可以根据相对丰度进行识别和分类,而 DRAGEN 软件仅报告主要群体。由于大多数开发用于分析 SARS-CoV-2 序列的软件旨在快速获得共识序列,因此样本中病毒群体的信息很少。在这里,我们提出了一种更快、更深层次的 SARS-CoV-2 监测方法,从 RT-qPCR 筛选到 NGS 分析。