急性下呼吸道感染病例呼吸道样本鼻病毒分离株基因型特征研究

Genotype characteristics of rhinovirus isolates from respiratory samples in patients with acute lower respiratory infection

目的人鼻病毒(human rhinovirus,HRV)是导致呼吸道感染的主要病原体之一。本研究分析急性下呼吸道感染病例样本中鼻病毒分离株基因型特征。方法急性下呼吸道感染病例肺泡灌洗液用核酸检测方法确定150例HRV阳性,样本接种H1-HeLa细胞,观察细胞病变,病毒纯化后用深度测序方法分析全基因组序列特征,用中和试验方法分析感染病例恢复期血清中和抗体水平。结果从21例病例样本中分离获得HRV病毒株。全基因组序列分析鉴定其为15个基因型,与参比株序列比较功能基因氨基酸具有多样性。3例HRV阳性病例恢复期血清中和抗体效价有4倍增高。结论成功分离获得HRV毒株,为进一步阐释鼻病毒在重症呼吸道感染的基础研究提供可用资源。

Objective Human rhinovirus(HRV)is one of the main pathogens causing respiratory infections.This study analyzed the genotype characteristics of rhinovirus isolates from bronchoalveolar lavage(BAL)of patients suffered with acute lower respiratory infections.Methods A total of 150 cases of HRV positive BAL samples were identified by nucleic acid test.The samples were inoculated on H1-HeLa cells and cytopathic effect were observed.The gene sequences of isolated HRV strains were obtained by using deep sequencing method after the virus purification.The convalescence sera of the patients were used in neutralization test to analyzed the neutralizing antibody levels.Results HRV strains were isolated from 21 samples.Phylogenic analysis and genome sequencing identified 15 genotypes.Compared with the reference sequences,the amino acid of the functional genes showed genetic diversity.Neutralization tests showed three HRV-positive cases had four-fold increase in neutralizing antibody titers in the convalescent sera.Conclusions The successful isolation of HRV strains will provide usable resources for further basic researches of the pathogenic role of viral rhinovirus in severe respiratory infections.