限制性内切酶分析快速检测严重急性呼吸系统综合征冠状病毒2型刺突蛋白484氨基酸突变的重要性

IF 0.1 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Investigacion clinica Pub Date : 2021-07-30 DOI:10.22209/IC.v62s2a02
R. Jaspe, Y. Sulbarn, C. Loureiro, Pierina DAngelo, L. Rodríguez, D. Garzaro, H. Rangel, F. Pujol
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引用次数: 2

摘要

导致新冠肺炎的新型冠状病毒SARS-CoV-2(VOCs)的变异毒株已在几个国家出现。刺突蛋白氨基酸484的突变特别重要,并与其中一些变体有关:E484K或E484Q。这些突变与逃避中和抗体有关。限制性内切酶分析被认为是一种快速检测这些突变的方法。2021年4月,对GISAID进行了搜索,以检测这两个突变在可用序列中的频率及其与其他谱系的关联。E484K存在于一些挥发性有机物中,已出现在其他几个谱系中,并经常在最近的病毒分离株中发现。Spike基因的小扩增子用两种酶消化:HpyAV和MseI。使用这两种酶可以检测484位的突变,并区分这三种情况:未突变和E484K或E484Q的存在。观察到与测序结果100%相关。所提出的方法可以筛选大量样本,可能有助于提供更多关于这些突变在全球范围内的流行率和流行病学的信息,以选择全基因组测序的候选者。
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Importance of mutations in amino acid 484 of the Spike protein of SARS-CoV-2: rapid detection by restriction enzyme analysis
Variants of Concern of SARS-CoV-2 (VOCs), the new coronavirus responsible for COVID-19, have emerged in several countries. Mutations in the amino acid 484 of the Spike protein are particularly important and associated with some of these variants: E484K or E484Q. These mutations have been associated with evasion to neutralizing antibodies. Restriction enzyme analysis is proposed as a rapid method to detect these mutations. A search on GISAID was performed in April 2021 to detect the frequency of these two mutations in the sequence available and their association with other lineages. E484K, present in some VOCs, has emerged in several other lineages and is frequently found in recent viral isolates. A small amplicon from the Spike gene was digested with two enzymes: HpyAV, and MseI. The use of these two enzymes allows the detection of mutations at position 484, and to differentiate between these three conditions: non-mutated, and the presence of E484K or E484Q. A 100% correlation was observed with sequencing results. The proposed methodology, which allows for the screening of a great number of samples, will probably help to provide more information on the prevalence and epidemiology of these mutations worldwide, to select the candidates for whole-genome sequencing.
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来源期刊
Investigacion clinica
Investigacion clinica MEDICINE, RESEARCH & EXPERIMENTAL-
CiteScore
0.20
自引率
50.00%
发文量
2
审稿时长
>12 weeks
期刊介绍: Estudios humanos, animales y de laboratorio relacionados con la investigación clínica y asuntos conexos.
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