婴幼儿疑似甲型H1N1重症病例及门诊流感样病例中鼻病毒的检测分析

目的 了解鼻病毒在婴幼儿疑似甲型H1N1重症病例及门诊流感样病例中的分布、分子进化特征，及与其他呼吸道病毒的合并感染情况。方法 样本有两种来源：疑似甲流重症病例样本246份为样本1组，门诊流感样病例样本68份为样本2组。扩增鼻病毒5’非编码区及VP2-VP4区，测序比对，构建进化树。对鼻病毒阳性样本检测其他常见呼吸道病毒。结果 样本1组中鼻病毒阳性率为8.54%(21/246)。样本2组中鼻病毒阳性率为16.2%（11/68）。样本1组和样本2组鼻病毒与其他呼吸道病毒的合并感染率分别为71.4%、9.09%，两者具有显著性差异（P<0.05）。对其中14份样本成功进行了测序，HRV-A、HRV-B、HRV-C基因型所占比例分别为64.3%、7.1%和28.6%。多序列比对分析表明鼻病毒3个基因型间的核苷酸序列一致性为55%～65%。结论 HRV在疑似甲型H1N1流感重症病例中主要是以合并感染为主。杭州地区婴幼儿疑似甲型H1N1流感重症病例及门诊流感样病例中鼻病毒感染主要是以HRV-A，HRV-C基因型为主，HRV各型别之间核苷酸变异较大。

Detection and Analysis of Rhinovirus in Children with Severe Influenza A (H1N1/2009)-like Infection and with Influenza-like Illness

The aim of the present research was to study the infection, prevalence and evolution of human rhinovirus in children with severe influenza A (H1N1/2009)-like infection and influenza-like illness, and to study the coinfection for HRV with the other respiratory viruses. Throat swab/endotracheal aspirates specimens were collected from 246 children with severe influenza A (H1N1/2009)-like infection (group 1), and throat swab specimens were collected from 68 children with influenza-like illness (group 2). Molecular tests were used to detect its 5’ noncoding region and VP2-VP4 of human rhinovirus. Nucleic acid sequence for VP2-VP4 was used for phylogenetic reconstruction. The co-infection for HRV with the other respiratory viruses was analysed. Results showed that in the 246 specimens of group 1, 21 specimens were HRV positive (8.54%). In the 68 specimens of group 2, 11 specimens were HRV positive (16.2%). The rate of co-infection for HRV with another viruses was 71.4% and 9.09% in group 1 and group 2, respectively, which had significant difference (P<0.05). Among the 14 specimens which were sequenced successfully, the rate for three HRV species, HRV-A, HRV-B, and the novel species of HRV-C, was 64.3%, 7.1%, and 28.6%, respectively. There were only 55％ to 65％ nucleotide acid identity among the three HRV genotypes. In conclusion, the main infection is coinfection of HRV in severe influenza A (H1N1/2009)-like illness. The main genotypes of human rhinovirus in children with severe influenza A (H1N1/2009) -like infection and with influenza-like illness are HRV-A and HRV-C in Hangzhou. The nucleotide acid of HRVs are quite variable.