HAV吕8株生长特性及残余活毒检测方法的研究

STUDY ON THE GROWTH CHARACTERISTICS OF HAV L8 AND DETECTING METHOD OF REMNANT LIVE VIRUS

[目的]对甲型肝炎病毒吕8株(HAV吕8)培养生长特性进行研究,确定其灭活后残余活毒检测方法的可靠性。[方法]通过一步生长曲线确定HAV吕8株的生长增殖高峰;将活病毒样品作2倍系列稀释后接种于细胞,研究不同接种量的最早检出时间;以统计学方法计算细胞培养方法检测HAV的灵敏性及可靠性;以病毒在细胞上的感染性为指标,经3次传代检测其有无感染性,以确证灭活疫苗残余活毒检测方法的可靠性。[结果]HAV吕8株细胞培养增殖高峰约为d20￣24;细胞培养方法检测甲肝病毒的灵敏性可达1.79CCID50、可靠检出甲肝病毒的最低量为10.23CCID50;凡达到ELISA的检出灵敏度的HAV接种样品,当培养增殖到d12时,均能被检出;当HAV含量达到512Eu/0.1ml时,不论是活病毒还是已灭活病毒,其吸附在细胞上的残余病毒量均能达到ELISA的检测灵敏度。[结论]将甲肝灭活疫苗样品接种细胞进行残余活毒检测,经3次传代,每次培养20d以上,该检测方法有较高的灵敏度,其检测结果是可靠的。

[Objective]To study the growth characteristics of HAV8 and the reliability of detecting method of remnant live virus of inactivated HAV8 vaccine.[Methods]One-step growth curve was used to determine the peak yield of HAV8.[Results]The peak replication of HAV L8 was from day 20 to 24 after inoculation,the detecting sensitivity by cell culture of HAV8 was 1.79 CCID50,and the lowest detectable quantity was 10.23 CCID50. The sample,which was in accord with the sensitivity of ELISA,can be detected 12 days after inoculation. When the content of either live virus or inactivated HAV8 was 512Eu/0.1ml,detecting sensitivity of remnant live virus in cell culture was in accord with that of ELISA.[Conclusion]If the sample of inactivated HAV L8 vaccine is passaged for three times by cell culture for over 20 days respectively,the remnant live virus can be detected. The method has highly sensitivity and reliability.