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高分辨率熔解曲线分析在 SARS-CoV-2 基因组监测中的潜在应用。

Potential use of high-resolution melting analyses for SARS-CoV-2 genomic surveillance.

机构信息

Serviço de Pesquisa em Doenças Infecciosas, Divisão de Ciência e Inovação, Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil.

Serviço de Pesquisa em Doenças Infecciosas, Divisão de Ciência e Inovação, Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil; Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.

出版信息

J Virol Methods. 2023 Jul;317:114742. doi: 10.1016/j.jviromet.2023.114742. Epub 2023 Apr 26.

Abstract

The pandemic caused by COVID-19 and the emergence of new variants of SARS-CoV-2 have generated clinical and epidemiological impacts on a global scale. The use of strategies for monitoring viral circulation and identifying mutations in genomic regions involved in host interaction are important measures to mitigate viral dissemination and reduce its likely complications on population health. In this context, the objective of this work was to explore the potential of high-resolution melting (HRM) analysis combined with one-step real-time reverse transcription PCR in a closed-tube system, as a fast and convenient method of screening for SARS-CoV-2 mutations with possible implications on host-pathogen interactions. The HRM analyses allowed the distinction of the Gamma, Zeta, Alpha, Delta, and Omicron variants against the predecessors (B.1.1.28, B.1.1.33) of occurrence in Brazil. It is concluded that the molecular tool standardized here has the potential to optimize the genomic surveillance of SARS-CoV-2, and could be adapted for genomic surveillance of other pathogens, due to its ability to detect, prior to sequencing, samples suggestive of new variants, selecting them more assertively and earlier for whole genome sequencing when compared to random screening.

摘要

由 COVID-19 引发的大流行和 SARS-CoV-2 的新变体的出现,在全球范围内产生了临床和流行病学的影响。监测病毒循环和识别宿主相互作用相关基因组区域突变的策略,是减轻病毒传播和降低其对人群健康可能产生的并发症的重要措施。在这种情况下,这项工作的目的是探索高分辨率熔解(HRM)分析与一步实时逆转录 PCR 在封闭管系统中的结合,作为一种快速便捷的方法,用于筛选可能影响宿主-病原体相互作用的 SARS-CoV-2 突变。HRM 分析能够区分伽马、泽塔、阿尔法、德尔塔和奥密克戎变体与在巴西发生的前体(B.1.1.28、B.1.1.33)。因此,这里标准化的分子工具具有优化 SARS-CoV-2 基因组监测的潜力,并且由于其能够在测序之前检测到提示新变体的样本,因此可以适应其他病原体的基因组监测,与随机筛选相比,该工具能够更有把握和更早地选择它们进行全基因组测序。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b670/10132831/301e8ab7bf0f/gr1_lrg.jpg

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