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在哈萨克斯坦使用牛津纳米孔技术通过全基因组测序对严重急性呼吸综合征冠状病毒2(SARS-CoV-2)进行高规模分析。

A high scale SARS-CoV-2 profiling by its whole-genome sequencing using Oxford Nanopore Technology in Kazakhstan.

作者信息

Kairov Ulykbek, Amanzhanova Amina, Karabayev Daniyar, Rakhimova Saule, Aitkulova Akbota, Samatkyzy Diana, Kalendar Ruslan, Kozhamkulov Ulan, Molkenov Askhat, Gabdulkayum Aidana, Sarbassov Dos, Akilzhanova Ainur

机构信息

Center for Life Sciences, National Laboratory Astana, Nazarbayev University, Nur-Sultan, Kazakhstan.

School of Sciences and Humanities, Nazarbayev University, Nur-Sultan, Kazakhstan.

出版信息

Front Genet. 2022 Sep 2;13:906318. doi: 10.3389/fgene.2022.906318. eCollection 2022.

Abstract

Severe acute respiratory syndrome (SARS-CoV-2) is responsible for the worldwide pandemic, COVID-19. The original viral whole-genome was sequenced by a high-throughput sequencing approach from the samples obtained from Wuhan, China. Real-time gene sequencing is the main parameter to manage viral outbreaks because it expands our understanding of virus proliferation, spread, and evolution. Whole-genome sequencing is critical for SARS-CoV-2 variant surveillance, the development of new vaccines and boosters, and the representation of epidemiological situations in the country. A significant increase in the number of COVID-19 cases confirmed in August 2021 in Kazakhstan facilitated a need to establish an effective and proficient system for further study of SARS-CoV-2 genetic variants and the development of future Kazakhstan's genomic surveillance program. The SARS-CoV-2 whole-genome was sequenced according to SARS-CoV-2 ARTIC protocol (EXP-MRT001) by Oxford Nanopore Technologies at the National Laboratory Astana, Kazakhstan to track viral variants circulating in the country. The 500 samples kindly provided by the Republican Diagnostic Center (UMC-NU) and private laboratory KDL "Olymp" were collected from individuals in Nur-Sultan city diagnosed with COVID-19 from August 2021 to May 2022 using real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR). All samples had a cycle threshold (Ct) value below 20 with an average Ct value of 17.03. The overall average value of sequencing depth coverage for samples is 244X. 341 whole-genome sequences that passed quality control were deposited in the Global initiative on sharing all influenza data (GISAID). The BA.1.1 ( = 189), BA.1 ( = 15), BA.2 ( = 3), BA.1.15 ( = 1), BA.1.17.2 ( = 1) omicron lineages, AY.122 ( = 119), B.1.617.2 ( = 8), AY.111 ( = 2), AY.126 ( = 1), AY.4 ( = 1) delta lineages, one sample B.1.1.7 ( = 1) belongs to alpha lineage, and one sample B.1.637 ( = 1) belongs to small sublineage were detected in this study. This is the first study of SARS-CoV-2 whole-genome sequencing by the ONT approach in Kazakhstan, which can be expanded for the investigation of other emerging viral or bacterial infections on the country level.

摘要

严重急性呼吸综合征冠状病毒2(SARS-CoV-2)引发了全球大流行疾病COVID-19。最初的病毒全基因组是通过高通量测序方法,对从中国武汉采集的样本进行测序得到的。实时基因测序是应对病毒爆发的主要参数,因为它能加深我们对病毒增殖、传播和进化的理解。全基因组测序对于SARS-CoV-2变异株监测、新疫苗和加强针的研发以及了解该国的疫情形势至关重要。2021年8月哈萨克斯坦确诊的COVID-19病例数量显著增加,因此有必要建立一个有效且高效的系统,以进一步研究SARS-CoV-2基因变异,并制定哈萨克斯坦未来的基因组监测计划。哈萨克斯坦国家实验室阿斯塔纳依据SARS-CoV-2 ARTIC方案(EXP-MRT001),采用牛津纳米孔技术对SARS-CoV-2全基因组进行测序,以追踪该国传播的病毒变异株。从2021年8月至2022年5月,通过实时逆转录定量聚合酶链反应(RT-qPCR),从努尔苏丹市确诊为COVID-19的个体中采集了500份样本,这些样本由共和国诊断中心(UMC-NU)和私人实验室KDL “Olymp” 慷慨提供。所有样本的循环阈值(Ct)值均低于20,平均Ct值为17.03。样本的测序深度覆盖总体平均值为244X。341条通过质量控制的全基因组序列已存入全球共享所有流感数据倡议组织(GISAID)。本研究检测到了BA.1.1(=189)、BA.1(=15)、BA.2(=3)、BA.1.15(=1)、BA.1.17.2(=1)奥密克戎谱系,AY.122(=119)、B.1.617.2(=8)、AY.111(=2)、AY.126(=1)、AY.4(=1)德尔塔谱系,一个样本B.1.1.7(=1)属于阿尔法谱系,以及一个样本B.1.637(=1)属于小分支。这是哈萨克斯坦首次采用ONT方法进行SARS-CoV-2全基因组测序研究,该研究可在国家层面扩展到对其他新出现的病毒或细菌感染的调查。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c79c/9479076/61ef02073cb2/fgene-13-906318-g001.jpg

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