College of Medicine, Taif University, Al-Taif 21944, Saudi Arabia.
Department of biology, College of Applied Sciences, Umm Al Qura University, Makkah 21955, Saudi Arabia.
Medicina (Kaunas). 2019 May 20;55(5):169. doi: 10.3390/medicina55050169.
The human respiratory syncytial virus (hRSV) is among the important respiratory pathogens affecting children. Genotype-specific attachment (G) gene sequencing is usually used to determine the virus genotype. The reliability of the fusion (F) gene vs. G gene genotype-specific sequencing was screened. Archival RNA from Saudi children who tested positive for hRSV-A were used. Samples were subjected to a conventional one-step RT-PCR for both F and G genes and direct gene sequencing of the amplicons using the same primer sets. Phylogeny and mutational analysis of the obtained sequences were conducted. The generic primer set succeeded to amplify target gene sequences. The phylogenetic tree based on partial F gene sequencing resulted in an efficient genotyping of hRSV-A strains equivalent to the partial G gene genotyping method. NA1, ON1, and GA5 genotypes were detected in the clinical samples. The latter was detected for the first time in Saudi Arabia. Different mutations in both conserved and escape-mutant domains were detected in both F and G. It was concluded that a partial F gene sequence can be used efficiently for hRSV-A genotyping.
人类呼吸道合胞病毒（hRSV）是影响儿童的重要呼吸道病原体之一。通常使用基因型特异性附着（G）基因测序来确定病毒基因型。本研究旨在筛选融合（F）基因与 G 基因基因型特异性测序的可靠性。使用沙特阿拉伯经检测 hRSV-A 阳性的儿童的存档 RNA。使用相同的引物对，对样本进行针对 F 和 G 基因的常规一步 RT-PCR 和扩增子的直接基因测序。对获得的序列进行系统发育和突变分析。通用引物成功扩增了目标基因序列。基于部分 F 基因测序的系统发育树可等效于部分 G 基因分型方法对 hRSV-A 株进行高效分型。在临床样本中检测到 NA1、ON1 和 GA5 基因型。后者在沙特阿拉伯首次被检测到。在 F 和 G 中均检测到保守和逃逸突变区的不同突变。结论是，部分 F 基因序列可有效用于 hRSV-A 基因分型。
