Department Experimental Medicine, University of Rome "Tor Vergata", 00133 Rome, Italy.
Via Luigi Einaudi, Guidonia Montecelio, 00012 Rome, Italy.
Viruses. 2023 Jan 26;15(2):353. doi: 10.3390/v15020353.
Currently, the reference method for identifying the presence of variants of SARS-CoV-2 is whole genome sequencing. Although it is less expensive than in the past, it is still time-consuming, and interpreting the results is difficult, requiring staff with specific skills who are not always available in diagnostic laboratories. The test presented in this study aimed to detect, using traditional real-time PCR, the presence of the main variants described for the spike protein of the SARS-CoV-2 genome. The primers and probes were designed to detect the main deletions that characterize the different variants. The amplification targets were deletions in the S gene: 25-27, 69-70, 241-243, and 157-158. In the gene, the deletion 3675-3677 was chosen. Some of these mutations can be considered specific variants, while others can be identified by the simultaneous presence of one or more deletions. We avoided using point mutations in order to improve the speed of the test. Our test can help clinical and medical microbiologists quickly recognize the presence of variants in biological samples (particularly nasopharyngeal swabs). The test can also be used to identify variants of the virus that could potentially be more diffusive as well as not responsive to the vaccine.
目前,鉴定 SARS-CoV-2 变异体存在的参考方法是全基因组测序。尽管它比过去便宜,但仍然耗时,并且结果解释困难,需要具有特定技能的人员,而这些人员在诊断实验室中并不总是能够提供。本研究中提出的测试旨在使用传统的实时 PCR 检测 SARS-CoV-2 基因组刺突蛋白中描述的主要变异体的存在。设计了引物和探针来检测表征不同变异体的主要缺失。扩增靶标是 S 基因中的缺失:25-27、69-70、241-243 和 157-158。在 基因中,选择了 3675-3677 的缺失。其中一些突变可以被认为是特定的变异体,而其他的则可以通过一个或多个缺失的同时存在来识别。我们避免使用点突变,以提高测试速度。我们的测试可以帮助临床和医学微生物学家快速识别生物样本(特别是鼻咽拭子)中变异体的存在。该测试还可用于识别可能更具扩散性且对疫苗无反应的病毒变异体。