Alsayed Ahmad R, Abed Anas, Abu-Samak Mahmoud, Alshammari Farhan, Alshammari Bushra
Department of Clinical Pharmacy and Therapeutics, Applied Science Private University, Amman 11931, Jordan.
Pharmacological and Diagnostic Research Centre, Faculty of Pharmacy, Al-Ahliyya Amman University, Amman 11931, Jordan.
J Clin Med. 2023 Jun 8;12(12):3909. doi: 10.3390/jcm12123909.
This research aims to determine acute bronchiolitis' causative virus(es) and establish a viable protocol to classify the Human Rhinovirus (HRV) species. During 2021-2022, we included children 1-24 months of age with acute bronchiolitis at risk for asthma. The nasopharyngeal samples were taken and subjected to a quantitative polymerase chain reaction (qPCR) in a viral panel. For HRV-positive samples, a high-throughput assay was applied, directing the VP4/VP2 and VP3/VP1 regions to confirm species. BLAST searching, phylogenetic analysis, and sequence divergence took place to identify the degree to which these regions were appropriate for identifying and differentiating HRV. HRV ranked second, following RSV, as the etiology of acute bronchiolitis in children. The conclusion of the investigation of all available data in this study distributed sequences into 7 HRV-A, 1 HRV-B, and 7 HRV-C types based on the VP4/VP2 and VP3/VP1 sequences. The nucleotide divergence between the clinical samples and the corresponding reference strains was lower in the VP4/VP2 region than in the VP3/VP1 region. The results demonstrated the potential utility of the VP4/VP2 region and the VP3/VP1 region for differentiating HRV genotypes. Confirmatory outcomes were yielded, indicating how nested and semi-nested PCR can establish practical ways to facilitate HRV sequencing and genotyping.
本研究旨在确定急性细支气管炎的致病病毒，并建立一种可行的方案来对人鼻病毒（HRV）进行分类。在2021年至2022年期间，我们纳入了1至24个月龄有哮喘风险的急性细支气管炎患儿。采集鼻咽样本，并在病毒检测板中进行定量聚合酶链反应（qPCR）。对于HRV阳性样本，应用高通量检测方法，针对VP4/VP2和VP3/VP1区域来确认病毒种类。进行BLAST搜索、系统发育分析和序列差异分析，以确定这些区域在识别和区分HRV方面的适用程度。在儿童急性细支气管炎的病因中，HRV仅次于呼吸道合胞病毒（RSV），排名第二。根据VP4/VP2和VP3/VP1序列，本研究对所有可用数据的调查结果将序列分为7种HRV-A、1种HRV-B和7种HRV-C型。临床样本与相应参考菌株之间的核苷酸差异在VP4/VP2区域低于VP3/VP1区域。结果证明了VP4/VP2区域和VP3/VP1区域在区分HRV基因型方面的潜在用途。得出了验证性结果，表明巢式和半巢式PCR如何能够建立切实可行的方法来促进HRV测序和基因分型。
