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目的研究从急性呼吸道感染(acute respiratory disease,ARD)患者咽拭样本中分离到的1株人腺病毒的基因型别。方法从辽宁省沈阳市采集ARD患者咽拭样本,用细胞培养方法分离病毒,对引起HEP-2细胞病变(CPE)的病毒用人腺病毒属通用引物进行聚合酶链反应(polymerase chain reaction,PCR)检测,并对阳性PCR产物进行测序分析;对病毒六邻体基因(HEXON)进行测序并与GenBank中其他人腺病毒参考毒株序列进行比较;用Mega 3.1软件、以邻位相连法构建人腺病毒属HEXON基因遗传进化树。结果从辽宁省沈阳市ARD患者咽拭样本中分离到1株病毒,人腺病毒属通用引物PCR检测阳性,阳性PCR产物测序结果与人腺病毒14型参考毒株序列完全一致;该腺病毒HEXON基因测序全长2838 bp,负责编码946个氨基酸,亦与人腺病毒14型参考毒株序列有100%的一致性;HEXON基因遗传进化树分析显示该腺病毒与人腺病毒14型参考毒株在系统发生树上处在同一进化分支上。结论辽宁省沈阳市ARD患者咽拭中分离到的腺病毒为人腺病毒14型,GenBank收录号为KC825052。
Objective To study the genotypes of a human adenovirus isolated from pharyngeal swab specimens from patients with acute respiratory disease (ARD). Methods A pharyngeal swab samples from patients with ARD were collected from Shenyang City, Liaoning Province. The virus was isolated by cell culture method. Polymerase chain reaction (PCR) was performed on the virus that caused HEP-2 cytopathic effect (CPE) (HEXON) was sequenced and compared with other human adenovirus reference strains in GenBank. Mega 3.1 software was used to construct the human gland HEXON gene is a phylogenetic tree. Results A virus was isolated from pharyngeal swab specimens of ARD patients in Shenyang, Liaoning Province. The positive PCR products of human adenoviruses were detected by PCR. The sequencing results of positive PCR products were completely consistent with the sequences of human adenovirus type 14 reference strains. The adenovirus HEXON The gene sequence was 2838 bp in length and was responsible for coding 946 amino acids and was 100% identical to that of human adenovirus type 14 reference strain. HEXON gene phylogenetic tree analysis showed that the adenovirus and human adenovirus type 14 reference strain The phylogenetic tree is on the same evolutionary branch. Conclusion The adenovirus isolated from pharyngeal swab of ARD patients in Shenyang of Liaoning Province is human adenovirus type 14, GenBank accession number is KC825052.