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[目的]分析人呼吸道合胞病毒(humanrespiratorysyncytialvirus,HRSV)经人二倍体KMB17细胞低温传代后培养特性的改变。[方法]将HRSVA2株病毒接种到人二倍体KMB17细胞进行低温培养传代,将不同代次的病毒经鼻腔接种豚鼠,于d7、d14分别取肺组织作病理切片检查、分离血清进行中和抗体检测,同时测定不同代次病毒的一步生长曲线及温度敏感性特征。[结果]豚鼠经鼻腔接种,接种原代病毒组d14处死时体重明显较对照组轻,低温第6代组和11代组与对照组接近,3组病毒均引发豚鼠间质性肺炎。低温第6代组和11代组与原代病毒相比,一步生长曲线未发生明显改变,增殖高峰约为9～12d。[结论]HRSV经鼻腔接种豚鼠能引起强烈的肺部感染,豚鼠可作为HRSV毒力评价的动物模型;HRSV在KMB17细胞上经低温传11代,未出现明显的减毒株特征,还需要进一步减毒。
[Objective] The research aimed to analyze the change of culture characteristics of human respiratory syncytial virus (HRSV) after passage of human diploid KMB17 cells at low temperature. [Method] HRSVA2 strain was inoculated into human diploid KMB17 cells for subculture at low temperature. Guinea pigs were inoculated with different generations of virus via nasal cavity. The lung tissues were taken for pathological examination on day 7 and day 14 respectively. The serum was separated for neutralizing antibody Detection, simultaneous determination of different generations of virus one-step growth curve and temperature-sensitive characteristics. [Results] Guinea pigs were inoculated intranasally and inoculated with the primary virus group, the body weight of mice killed at d14 was significantly lower than that of the control group. The 6th and 11th generation hypothermia groups were close to the control group, and all three groups of viruses caused guinea pig interstitial pneumonia. Compared with the primary virus, the one-step growth curve did not change significantly in the 6th and 11th generation hypothermia groups, and the peak of proliferation was about 9-12 days. [Conclusion] HRSV intranasal inoculation of guinea pigs can cause strong lung infection, guinea pigs can be used as animal models of HRSV virulence assessment; HRSV in KMB17 cells after cryogenic transmission of 11 generations, did not appear obvious attenuated strains characteristics, but also further Attenuated.