Usserbayev Bekbolat, Sultankulova Kulyaisan T, Burashev Yerbol, Melisbek Aibarys, Shirinbekov Meirzhan, Myrzakhmetova Balzhan S, Zhunushov Asankadir, Smekenov Izat, Kerimbaev Aslan, Nurabaev Sergazy, Chervyakova Olga, Kozhabergenov Nurlan, Kutumbetov Lesbek B
Research Institute for Biological Safety Problems, National Holding QazBioPharm, LLP, Guardeyskiy uts 080409, Kazakhstan.
Institute of Biotechnology, National Academy of Science of Kyrgyzstan, Bishkek 720071, Kyrgyzstan.
Viruses. 2025 Mar 14;17(3):415. doi: 10.3390/v17030415.
Prompt determination of the etiological agent is important in an outbreak of pathogens with pandemic potential, particularly for dangerous infectious diseases. Molecular genetic methods allow for arriving at an accurate diagnosis, employing timely preventive measures, and controlling the spread of the disease-causing agent. In this study, whole-genome sequencing of three SARS-CoV-2 strains was performed using the Sanger method, which provides high accuracy in determining nucleotide sequences and avoids errors associated with multiple DNA amplification. Complete nucleotide sequences of samples, KAZ/Britain/2021, KAZ/B1.1/2021, and KAZ/Delta020/2021 were obtained, with sizes of 29.751 bp, 29.815 bp, and 29.840 bp, respectively. According to the COVID-19 Genome Annotator, 127 mutations were detected in the studied samples compared to the reference strain. The strain KAZ/Britain/2021 contained 3 deletions, 7 synonymous mutations, and 27 non-synonymous mutations, the second strain KAZ/B1.1/2021 contained 1 deletion, 5 synonymous mutations, and 31 non-synonymous mutations, and the third strain KAZ/Delta020/2021 contained 1 deletion, 5 synonymous mutations, and 37 non-synonymous mutations, respectively. The variations C241T, F106F, P314L, and D614G found in the 5' UTR, , and regions were common to all three studied samples, respectively. According to PROVEAN data, the loss-of-function mutations identified in strains KAZ/Britain/2021, KAZ/B1.1/2021, and KAZ/Delta020/2021 include 5 mutations (P218L, T716I, W149L, R52I, and Y73C), 2 mutations (S813I and Q992H), and 8 mutations (P77L, L452R, I82T, P45L, V82A, F120L, F120L, and R203M), respectively. Phylogenetic analysis showed that the strains studied (KAZ/Britain/2021, KAZ/B1.1/2021, and KAZ/Delta020/2021) belong to different SARS-CoV-2 lineages, which are closely related to samples from Germany (OU141323.1 and OU365922.1), Mexico (OK432605.1), and again Germany (OV375251.1 and OU375174.1), respectively. The nucleotide sequences of the studied SARS-CoV-2 virus strains were registered in the Genbank database with the accession numbers: ON692539.1, OP684305, and OQ561548.1.
对于具有大流行潜力的病原体爆发,尤其是危险的传染病,迅速确定病原体至关重要。分子遗传学方法有助于做出准确诊断、及时采取预防措施并控制致病因子的传播。在本研究中,使用桑格法对三株严重急性呼吸综合征冠状病毒2(SARS-CoV-2)毒株进行了全基因组测序,该方法在确定核苷酸序列方面具有很高的准确性,并且避免了与多次DNA扩增相关的错误。获得了样本KAZ/英国/2021、KAZ/B1.1/2021和KAZ/Delta020/2021的完整核苷酸序列,其大小分别为29,751 bp、29,815 bp和29,840 bp。根据2019冠状病毒病基因组注释器,与参考毒株相比,在所研究的样本中检测到127个突变。毒株KAZ/英国/2021包含3个缺失、7个同义突变和27个非同义突变,第二株毒株KAZ/B1.1/2021包含1个缺失、5个同义突变和31个非同义突变,第三株毒株KAZ/Delta020/2021分别包含1个缺失、5个同义突变和37个非同义突变。在5'非翻译区、……和……区域发现的C241T、F106F、P314L和D614G变异分别在所有三个研究样本中都存在。根据蛋白质变异效应分析(PROVEAN)数据,在毒株KAZ/英国/2021、KAZ/B1.1/2021和KAZ/Delta020/2021中鉴定出的功能丧失突变分别包括5个突变(P218L、T716I、W149L、R52I和Y73C)、2个突变(S813I和Q992H)和8个突变(P77L、L452R、I82T、P45L、V82A、F120L、F120L和R203M)。系统发育分析表明,所研究的毒株(KAZ/英国/2021、KAZ/B1.1/2021和KAZ/Delta020/2021)属于不同的SARS-CoV-2谱系,它们分别与来自德国的样本(OU141323.1和OU365922.1)、墨西哥的样本(OK432605.1)以及再次来自德国的样本(OV375251.1和OU375174.1)密切相关。所研究的SARS-CoV-2病毒毒株的核苷酸序列已在Genbank数据库中注册,登录号分别为:ON692539.1、OP684305和OQ561548.1。