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用 RT-qPCR 筛查关注的 SARS-CoV-2 变体面临的挑战:一种变体可以隐藏另一种变体。

The challenge of screening SARS-CoV-2 variants of concern with RT-qPCR: One variant can hide another.

机构信息

Laboratory Medicine, Iris Hospitals South, 63 rue Jean Paquot, 1050 Brussels, Belgium.

Laboratory Medicine, Iris Hospitals South, 63 rue Jean Paquot, 1050 Brussels, Belgium.

出版信息

J Virol Methods. 2021 Nov;297:114248. doi: 10.1016/j.jviromet.2021.114248. Epub 2021 Jul 29.

Abstract

INTRODUCTION

Following the emergence of SARS-CoV-2 variants of concern (VOCs) worldwide, it is important to monitor local epidemiology to better understand the occurrence of clusters, reinfections, or infection after vaccination. Detecting mutations by specific RT-qPCR is a rapid and affordable alternative to sequencing. However, care must be taken to ensure that the techniques used are up-to-date and adapted to the variants circulating in the studied population.

MATERIAL AND METHODS

All samples tested positive for SARS-CoV-2 were screened for detection of mutations of the spike protein using the Novaplex™ SARS-CoV-2 Variants I Assay from week 11 of 2021. Target sought were deletion H69/V70 and mutations N501Y and E484K. From week 18 we used in addition the new Novaplex™ SARS-CoV-2 Variants II Assay for samples with no targets found with the Variants I assay or with the mutation E484K alone, in order to screen the mutations L452R, K417N/T and W152C.

RESULTS

Between weeks 11 and 25, 2239 positive samples out of 54,317 were tested with the Variants I Assay. Between weeks 18 and 25, 94 samples met the criteria for being tested with the Variants II Assay. Of these, 47 had the L452R mutation without the W152C mutation, typical in the B.1.617 variant. At week 25, this profile was found in 45.5 % of the samples and was the most frequent.

CONCLUSION

According to our observations, variant B.1.617 has become predominant in our institution and most probably in our region. In the absence of the use of the Variants II Assay, they would have been considered wild.

摘要

简介

随着 SARS-CoV-2 变异株(VOCs)在全球范围内的出现,监测当地流行病学以更好地了解聚集性、再感染或接种疫苗后的感染情况非常重要。通过特定的 RT-qPCR 检测突变是一种快速且经济实惠的替代测序方法。然而,必须注意确保使用的技术是最新的,并适应研究人群中流行的变体。

材料和方法

从 2021 年第 11 周开始,使用 Novaplex™ SARS-CoV-2 变体 I 检测试剂盒对所有检测呈 SARS-CoV-2 阳性的样本进行筛查,以检测刺突蛋白的突变。目标是检测缺失 H69/V70 和突变 N501Y 和 E484K。从第 18 周开始,我们还使用了新的 Novaplex™ SARS-CoV-2 变体 II 检测试剂盒,用于那些在变体 I 检测试剂盒或仅存在 E484K 突变的样本中未发现目标的样本,以筛选 L452R、K417N/T 和 W152C 突变。

结果

在第 11 周至第 25 周期间,对 54317 个样本中的 2239 个阳性样本进行了变体 I 检测。在第 18 周至第 25 周期间,有 94 个样本符合变体 II 检测的标准。其中,47 个样本具有 L452R 突变但没有 W152C 突变,这是 B.1.617 变体的典型特征。在第 25 周,这种情况在 45.5%的样本中发现,是最常见的。

结论

根据我们的观察,变体 B.1.617 已成为我们机构和我们地区的主要变体。如果不使用变体 II 检测试剂盒,它们将被视为野生型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c777/8320429/cd8004c14f81/gr1_lrg.jpg

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