人呼吸道合胞病毒A_2株在不同细胞中的培养及其影响因素

目的在不同细胞中培养人呼吸道合胞病毒(HRSV),并分析其影响因素,为病毒的大量增殖创造条件。方法将HRSVA2株分别接种至人二倍体KMB17细胞、Vero细胞、人喉癌Hep-2细胞及HeLa细胞进行培养,采用微量细胞病变法检测病毒的感染性滴度,并进行间接免疫荧光试验及合胞体观察。检测吸附时间、维持液pH值和培养温度对病毒增殖的影响。结果4种细胞中,Vero细胞对HRSV最敏感,病变早、病毒感染性滴度可达6.83lgCCID50/ml;KMB17细胞相对敏感性不高,但仍能良好增殖,病毒感染性滴度达6.13lgCCID50/ml;Hep-2细胞及HeLa细胞介于Vero细胞和KMB17细胞之间,病毒感染性滴度也达到6.83lgCCID50/ml。间接免疫荧光试验显示病毒在细胞浆内增殖。吸附时间对HRSV的增殖无明显影响;维持液的pH值对HRSV的增殖及病变特点均有一定的影响,在pH7.0～7.2时可快速引起细胞产生病变,并能达到最高感染性滴度;HRSV在37℃及32℃下培养均能良好地增殖。结论HRSVA2株在KMB17、Vero、Hep-2及HeLa细胞内均可增殖,但敏感性有一定差异,且对培养条件有一定的要求。

Culture of Human Respiratory Syncytial Virus A2 Strain in Various Cells and Its Influencing Factors

Objective To culture human respiratory syncytial virus(HRSV)A2 strain in various cells and analyze its influencing factors to provide condition for the propagation of HRSV in a large quantity.Methods HRSV A2 strain was cultured in human diploid KMB17, Vero, human laryngeal carcinoma Hep-2 and HeLa cells respectively, then determined for infectious titer by micro-CPE method, subjected to indirect IFA and observed for synplasm.The effect of time for adsorption, pH value of maintenance medium and temperature for culture on propagation of HRSV were analyzed.Results Of the 4 kinds of cells, Vero cells showed the highest senstivity to HRSV, in which CPE appeared early and the infectious titer of HRSV reached 6.83 lgCCID50 / ml.The sensitivities of Hep-2 and HeLa cells to HRSV were between those of Vero and KMB17 cells, in which the infectious titers also reached 6.83 lgCCID50 / ml.The sensitivity of KMB17 cells to HRSV was relative low, in which the infectious titer was 6.13 lgCCID50 / ml.Indirect IFA showed propagation of HRSV in cytoplasm.The time for adsorption showed no significant effect on HRSV propagation.The pH value of maintenance medium showed a certain effect on HRSV propagation and CPE.At pH 7.0～7.2, CPE appeared rapidly, and the in-fectious titer of HRSV reached a peak value.HRSV was propagated well at both 37 and 32℃.Conclusion HRSV A2 strain may be propagated in KMB17, Vero, Hep-2 and HeLa cells.However, the sensitivities of the cells to HRSV were different, and the culture condition showed a certain effect on HRSV propagation.