Aslan Ferhat Gürkan, Yalçin Süleyman, Kazci Saliha, Dinç Bedia, Korukluoğlu Gülay
Ankara Bilkent City Hospital, Clinic of Medical Microbiology, Ankara, Türkiye.
Türkiye Directorate General of Public Health, Department of Microbiology Reference and Biological Products, Ankara, Türkiye.
Mikrobiyol Bul. 2024 Oct;58(4):448-460. doi: 10.5578/mb.20249665.
The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) virus has mutated at a high rate since the beginning of the pandemic, leading to the formation of different variants. Alpha, Beta, Gamma, Delta and Omicron have emerged as concerning variants identified by the World Health Organization (WHO). The Omicron variant and its sublineages became dominant worldwide in 2022. EG.5, a sublineage of Omicron, was associated with increased cases recently. In this study, we aimed to investigate the relationship between SARS-CoV-2 variants and the severity of the disease by using the whole genome sequence analysis method in patient samples diagnosed with coronavirus diseases 2019 (COVID-19) in our hospital and to contribute to SARS-CoV-2 surveillance in our country, in the period until the end of 2023, after the announcement of the EG.5 variant. The study included 68 patient samples that were found to be SARS-CoV-2 positive by polymerase chain reaction. Nasopharyngeal/oropharyngeal swab samples obtained from the patients were analysed by syndromic multiplex viral polymerase chain reaction panel. Whole genome sequence analysis was performed on SARS-CoV-2 positive patient samples with high viral load. Barcoded sequencing library was prepared and extracted DNA was sequenced on the Illumina next generation sequencing platform using COVIDSeq test kits (Illumina, USA). The library was loaded into a NextSeq 500/550 high output reagent cartridge v2 75 cycle, NextSeq 500/550 high output flow cell on the NextSeq 550 instrument (Illumina, USA). The DRAGEN COVID lineage programme available in the core domain was used to analyse the sequence data. The resulting combined fasta files were loaded into Nexclade and Pangolin COVID-19 Lineage Assigner programmes and class assignment, mutation calls and sequence quality checks were performed. Statistical evaluation was performed using Statistical Package for Social Sciences (Windows v20.0, Armonk, NY) programme. Sequence results of a total of 68 patients whose SARS-CoV-2 positivity was confirmed in our laboratory and whose samples were suitable for sequence study were analysed with clinical data. While EG.5 variant was detected in 15 (22.1%) of the samples included in the study, different sub-variants other than EG.5 were detected in 53 (77.9%) samples. The first three most frequently detected variants were XBB.1.16, EG.5.1 and FL.1.5.1, respectively. The subtypes of the EG.5 variant were determined as EG.5.1.1, EG.5.1.3, EG.5.1.6 and EG.5.1. Of the patients included in the study, 20 (29.4%) were hospitalised and followed up, 14 of whom had moderate to severe clinical conditions. Four hospitalised patients with comorbidities at an advanced age resulted in exitus. In the statistical evaluation, no significant difference was found between EG.5 and nonEG.5 Omicron variants in terms of age group, clinical symptoms and disease severity. It is recommended to continue genomic surveillance for the timely identification of SARS-CoV-2 and changes in the clinical spectrum and implementation of appropriate countermeasures.
自疫情开始以来,严重急性呼吸综合征冠状病毒2(SARS-CoV-2)病毒一直以高频率发生变异,导致形成了不同的变体。阿尔法、贝塔、伽马、德尔塔和奥密克戎已成为世界卫生组织(WHO)确定的令人担忧的变体。奥密克戎变体及其亚谱系在2022年成为全球主导。奥密克戎的一个亚谱系EG.5最近与病例增加有关。在本研究中,我们旨在通过对我院诊断为2019冠状病毒病(COVID-19)的患者样本使用全基因组序列分析方法,研究SARS-CoV-2变体与疾病严重程度之间的关系,并在EG.5变体公布后至2023年底期间为我国的SARS-CoV-2监测做出贡献。该研究纳入了68份经聚合酶链反应检测发现SARS-CoV-2呈阳性的患者样本。从患者处采集的鼻咽/口咽拭子样本通过症状性多重病毒聚合酶链反应检测板进行分析。对病毒载量高的SARS-CoV-2阳性患者样本进行全基因组序列分析。制备带条形码的测序文库,并使用COVIDSeq检测试剂盒(美国Illumina公司)在Illumina下一代测序平台上对提取的DNA进行测序。该文库被加载到NextSeq 500/550高输出试剂盒v2 75循环、NextSeq 500/550高输出流动槽上的NextSeq 550仪器(美国Illumina公司)中。使用核心域中可用的DRAGEN COVID谱系程序分析序列数据。将得到的合并fasta文件加载到Nexclade和穿山甲COVID-19谱系分配程序中,并进行类别分配、突变检测和序列质量检查。使用社会科学统计软件包(Windows v20.0,纽约州阿蒙克)程序进行统计评估。对我院实验室确认SARS-CoV-2阳性且样本适合序列研究的68名患者的序列结果与临床数据进行了分析。在纳入研究的样本中,15份(22.1%)检测到EG.5变体,53份(77.9%)样本检测到EG.5以外的不同亚变体。检测到的前三个最常见变体分别是XBB.1.16、EG.5.1和FL.1.5.1。EG.5变体的亚型被确定为EG.5.1.1、EG.5.1.3、EG.5.1.6和EG.5.1。纳入研究的患者中有20名(29.4%)住院并接受随访,其中14名有中度至重度临床状况。4名患有合并症的老年住院患者死亡。在统计评估中,EG.5与非EG.5奥密克戎变体在年龄组、临床症状和疾病严重程度方面未发现显著差异。建议继续进行基因组监测,以便及时识别SARS-CoV-2及其临床谱的变化并实施适当的应对措施。