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与严重急性呼吸综合征冠状病毒2(SARS-CoV-2)基因组变异相关的突变评估:逆转录定量聚合酶链反应(RT-qPCR)作为监测2021年智利已知流行变异株的快速且经济实惠的工具

Assessment of Mutations Associated With Genomic Variants of SARS-CoV-2: RT-qPCR as a Rapid and Affordable Tool to Monitoring Known Circulating Variants in Chile, 2021.

作者信息

Angulo Jenniffer, Martinez-Valdebenito Constanza, Pardo-Roa Catalina, Almonacid Leonardo I, Fuentes-Luppichini Eugenia, Contreras Ana Maria, Maldonado Constanza, Le Corre Nicole, Melo Francisco, Medina Rafael A, Ferrés Marcela

机构信息

Departamento de Enfermedades Infeciosas e Inmmunologia Pediatricas, Escuela de Medicina, Pontificia Universidad Católica de Chile, Santiago, Chile.

Infectious Disease and Molecular Virology Laboratory, Red Salud UC-Christus, Santiago, Chile.

出版信息

Front Med (Lausanne). 2022 Feb 25;9:841073. doi: 10.3389/fmed.2022.841073. eCollection 2022.

Abstract

Since the first report of SARS-CoV-2 infection in humans, the virus has mutated to develop new viral variants with higher infection rates and more resistance to neutralization by antibodies elicited after natural SARS-CoV-2 infection or by vaccines. Therefore, rapid identification of viral variants circulating in the population is crucial for epidemiological assessment and efforts to contain the resurgence of the pandemic. Between January and November 2021, we performed a large variant RT-qPCR-based screening of mutations in the spike protein of 1851 SARS-CoV-2-positive samples derived from outpatients from the UC-Christus Health Network in Chile. In a portion of samples ( = 636), we validated our RT-qPCR-pipeline by WGS, obtaining a 99.2% concordance. Our results indicate that from January to March 2021 there was a dominance of non-identifiable variants by the RT-qPCR-based screening; however, throughout WGS we were able to identify the Lambda (C.37) variant of interest (VOI). From March to July, we observed the rapid emergence of mutations associated with the Gamma variant (P.1), which was quickly replaced by the appearance of a combination of samples harboring mutations associated with the Delta variant (B.1.617.2), which predominated until the end of the study. Our results highlight the applicability of cost-effective RT-qPCR-based screening of mutations associated with known variants of concern (VOC), VOI and variants under monitoring (VUM) of SARS-CoV-2, being a rapid and reliable tool that complements WGS-based surveillance.

摘要

自首次报告人类感染严重急性呼吸综合征冠状病毒2(SARS-CoV-2)以来,该病毒已发生变异,产生了新的病毒变体,这些变体具有更高的感染率,并且对自然感染SARS-CoV-2后或接种疫苗后产生的抗体中和作用具有更强的抵抗力。因此,快速识别在人群中传播的病毒变体对于流行病学评估和遏制疫情复发的努力至关重要。2021年1月至11月,我们对来自智利UC-Christus健康网络门诊患者的1851份SARS-CoV-2阳性样本的刺突蛋白突变进行了基于逆转录定量聚合酶链反应(RT-qPCR)的大规模变体筛查。在一部分样本(n = 636)中,我们通过全基因组测序(WGS)验证了我们的RT-qPCR流程,一致性达到99.2%。我们的结果表明,2021年1月至3月,基于RT-qPCR的筛查无法识别出优势变体;然而,通过WGS我们能够识别出感兴趣的拉姆达(C.37)变体(VOI)。从3月到7月,我们观察到与伽马变体(P.1)相关的突变迅速出现,随后很快被携带与德尔塔变体(B.1.617.2)相关突变的样本组合所取代,德尔塔变体在研究结束前一直占主导地位。我们的结果突出了基于RT-qPCR的具有成本效益的筛查与SARS-CoV-2已知关注变体(VOC)、感兴趣变体(VOI)和监测变体(VUM)相关突变的适用性,它是一种快速可靠的工具,可补充基于WGS的监测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2372/8914012/73297dc66ede/fmed-09-841073-g0001.jpg

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