摘要
目的探讨不同细胞对柯萨奇病毒A组10型(coxsackievirus A10,CV-A10)毒株的敏感性和滴度检测影响因素,为临床样品分离、病毒培养及保存使用提供基础数据。方法10株已在非洲绿猴肾(African green monkey kidney,Vero)细胞分离并培养2代的CV-A10毒株,检测其在Vero细胞、人胚肺二倍体细胞(human embryonic lung diploid cell,KMB17)、人横纹肌肉瘤(human rhabdomyosarcoma,RD)细胞及人喉癌细胞(human laryngeal carcinoma cell,HeP-2)上的感染性滴度,并对毒株进行蚀斑纯化和冻融处理。结果CV-A10毒株均能适应Vero细胞、KMB17细胞及RD细胞,不能适应HeP-2细胞。RD细胞上第1天即开始病变,第4天达到增值高峰,感染性滴度最高,达到7.75 lgCCID50/ml;KMB17细胞上病变最慢,第3天发生病变,第5天达到增值高峰,滴度为4.85 lgCCID50/ml。毒株在37℃放置3 d感染性滴度明显下降,4℃放置3 d,-30℃和-80℃放置30 d感染性滴度均无影响。结论RD细胞是CV-A10毒株最敏感的细胞,HeP-2细胞不敏感。CV-A10毒株反复冻融次数不宜超过5次,保存温度越低越好并避免4℃和常温下长时间放置。
Objective To study the factors influencing the sensitivity and titer detection of different cells to coxsackievirus A10(CV-A10).Methods Ten CV-A10 strains isolated and cultured in African green monkey kidney cells(Vero)for 2 generations were detected on Vero,human embryonic lung diploid cell(KMB17),human rhabdomyosarcoma(RD)and human laryngeal carcinoma cell(HeP-2).Virus strain was subjected to plaque purification and freeze-thaw treatment.Results All CV-A10 strains could adapt to Vero,KMB17 and RD cells,but could not adapt to HeP-2 cells.Viral strains showed the fastest time in RD cells to start lesions on the 1 st day and reached peak on the 4 th day,with the highest infectious titer at 7.75 lgCCID50/ml;the slowest lesions on KMB17 cells that occurred on the 3 rd day and peaked on the 5 th day with titer at 4.85 lgCCID50/ml.The infectious titer of the virus strain was significantly reduced after being placed at 37℃for 3 d,and the infectious titer was not affected by being placed at 4℃for 3 d,and being kept at-30℃and-80℃for 30 d.Conclusions RD cells are the most sensitive cells to CV-A10,and HeP-2 cells are not sensitive.The frequency of repeated freezing and thawing of the virus should not exceed 5 times.The lowest storage temperature of-80℃is better.The virus sample should be avoided for prolonged storage at 4℃and normal temperature.
作者
高微捷
岳磊
杨婷
宋霞
李华
谢天宏
何鑫
卫星辰
谢忠平
GAO Wei-jie;YUE Lei;YANG Ting;SONG Xia;LI Hua;XIE Tian-hong;HE Xin;WEI Xing-chen;XIE Zhong-ping(Key Laboratory for Vaccine Research and Development of Major Infectious Diseases in Yunnan Province,Institute of Medical Biology,Chinese Academy of Medical Sciences,Kunming,Yunnan 650118,China)
出处
《中国病毒病杂志》
CAS
2020年第4期267-271,共5页
Chinese Journal of Viral Diseases
基金
云南省科技计划项目(2015BC008)
中国医学科学院医学与健康科技创新工程项目(2016-I2M-1-019)。
