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本文引用的文献

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SARS-CoV-2 Variants and Clinical Outcomes: A Systematic Review.严重急性呼吸综合征冠状病毒2变种与临床结局:一项系统评价。
Life (Basel). 2022 Jan 25;12(2):170. doi: 10.3390/life12020170.
2
The Disease Severity and Clinical Outcomes of the SARS-CoV-2 Variants of Concern.关注的 SARS-CoV-2 变异株的疾病严重程度和临床结局。
Front Public Health. 2021 Nov 30;9:775224. doi: 10.3389/fpubh.2021.775224. eCollection 2021.
3
A benchmarking of human mitochondrial DNA haplogroup classifiers from whole-genome and whole-exome sequence data.全基因组和全外显子组序列数据中人线粒体 DNA 单倍群分类器的基准测试。
Sci Rep. 2021 Oct 15;11(1):20510. doi: 10.1038/s41598-021-99895-5.
4
Distinguishing features of current COVID-19 vaccines: knowns and unknowns of antigen presentation and modes of action.当前新冠疫苗的显著特征:抗原呈递和作用方式的已知与未知
NPJ Vaccines. 2021 Aug 16;6(1):104. doi: 10.1038/s41541-021-00369-6.
5
Characterization of SARS-CoV-2 different variants and related morbidity and mortality: a systematic review.SARS-CoV-2 不同变体的特征及其相关发病率和死亡率:系统评价。
Eur J Med Res. 2021 Jun 8;26(1):51. doi: 10.1186/s40001-021-00524-8.
6
The Spike D614G mutation increases SARS-CoV-2 infection of multiple human cell types.刺突 D614G 突变增加了 SARS-CoV-2 对多种人类细胞类型的感染。
Elife. 2021 Feb 11;10:e65365. doi: 10.7554/eLife.65365.
7
SARS-CoV-2 spike-protein D614G mutation increases virion spike density and infectivity.SARS-CoV-2 刺突蛋白 D614G 突变增加了病毒粒子刺突密度和感染力。
Nat Commun. 2020 Nov 26;11(1):6013. doi: 10.1038/s41467-020-19808-4.
8
Spike mutation D614G alters SARS-CoV-2 fitness.刺突突变 D614G 改变了 SARS-CoV-2 的适应性。
Nature. 2021 Apr;592(7852):116-121. doi: 10.1038/s41586-020-2895-3. Epub 2020 Oct 26.
9
Evolutionary and structural analyses of SARS-CoV-2 D614G spike protein mutation now documented worldwide.SARS-CoV-2 D614G 刺突蛋白突变的进化和结构分析现已在全球范围内记录。
Sci Rep. 2020 Aug 20;10(1):14031. doi: 10.1038/s41598-020-70827-z.
10
Identification of Common Deletions in the Spike Protein of Severe Acute Respiratory Syndrome Coronavirus 2.鉴定严重急性呼吸综合征冠状病毒 2 的刺突蛋白中的常见缺失。
J Virol. 2020 Aug 17;94(17). doi: 10.1128/JVI.00790-20.

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.

DOI:10.3390/ijms23063143
PMID:35328562
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8950365/
Abstract

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.

摘要

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。