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用于快速筛选 SARS-CoV-2 中 B.1.617 突变的变异特异性 RT-qPCR。

Variant-specific RT-qPCR for rapid screening of B.1.617 mutations in SARS-CoV-2.

机构信息

Genetic Engineering Department, Libyan Biotechnology Research Center, Tripoli, Libya.

Department of Microbiology, Libyan Biotechnology Research Centre, Tripoli, Libya.

出版信息

Libyan J Med. 2022 Dec;17(1):2121252. doi: 10.1080/19932820.2022.2121252.

Abstract

The continuous emergence of new SARS-CoV-2 variants required rapid and reliable diagnostic methods for early detection and monitoring of the spread of the virus, especially in low-resource countries where whole genome sequencing is not available. We aimed to evaluate and compare the performance of two different RT-qPCR screening assays for the detection of B.1.617 lineage mutations. A total of 85 SARS-CoV-2 positive samples were collected between 9 August and 10 September 2021 and screened by two mutation-specific RT-qPCR assays for simultaneous detection of B.1.617.1 and B.1.617.2 lineage mutations. VIASURE Variant II PCR assay identified 2 Delta variant-specific mutations (L452R, and P681 R) in 80% of tested samples, while the PKamp™ Variant Detect™ assay was only able to detect one Delta variant specific mutation (L452R) in 75% of tested samples. This is the first report to show the Delta variant as the cause of the third wave in Libya. The use of multiplex RT-qPCR assays has allowed the identification of new variants for rapid screening. However, RT-qPCR results should be confirmed by whole genome sequencing of SARS-COV-2.

摘要

不断出现的新型 SARS-CoV-2 变体需要快速可靠的诊断方法,以便早期检测和监测病毒的传播,特别是在无法进行全基因组测序的资源有限的国家。我们旨在评估和比较两种不同的 RT-qPCR 筛选检测方法在检测 B.1.617 谱系突变方面的性能。总共收集了 2021 年 8 月 9 日至 9 月 10 日之间的 85 个 SARS-CoV-2 阳性样本,并通过两种突变特异性 RT-qPCR 检测方法对 B.1.617.1 和 B.1.617.2 谱系突变进行了同时检测。VIASURE Variant II PCR 检测法在 80%的测试样本中发现了 2 种 Delta 变体特异性突变(L452R 和 P681R),而 PKamp™ Variant Detect™ 检测法仅在 75%的测试样本中检测到 1 种 Delta 变体特异性突变(L452R)。这是第一份显示德尔塔变异株是利比亚第三波疫情的原因的报告。多重 RT-qPCR 检测法的使用可以快速筛选鉴定出新的变体。然而,RT-qPCR 结果应通过对 SARS-COV-2 的全基因组测序进行确认。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c6c/9467536/85f9759f34b0/ZLJM_A_2121252_F0001_OC.jpg

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