摘要
为了研制高致病性猪繁殖与呼吸综合征病毒(PRRSV)国家核酸标准物质,将PRRSV美洲变异株代表毒株PRRSV JXA1接种Marc145细胞,出现细胞病变后,反复冻融并收集病毒液,并灭活分装。制备后进行均匀性、稳定性评估、数字PCR法定值、不确定度分析和临床试用。结果表明,制备的标准物质的定值为:(1.13±0.18)×10~4拷贝/μL,样品均匀;-20℃可稳定保存12个月以上、4℃可稳定2个月以上。经全国标准物质管理委员会的专家评审,该制备物达到了国家二级标准物质要求,可以作为核酸扩增检测PRRVS的核酸标准物质。
To prepare the national reference material of HP-PRRSV for nucleic acid amplification technology,Marc145 cell lines were infected by HP-PRRSV(JXA1).After the cytopathic effect was observed,the viral fluid was harvested by freezing and thawing repeatedly,inactivated and sub-packed.The homogeneity and stability of the prepared reference materials were tested.Then,the prepared reference materials were valued using digital PCR and the clinical applicability was also evaluated.The results indicated that the value of the reference material was(1.13±0.18)×10~4 copies/μL.The prepared materials could be stable at-20℃for at least 12 months,at 4℃for more than 2 months.The prepared material of PRRSV passed the review of national administrative committee for CRM’s,and reached the requirements of the national secondary reference.This reference material can be used as the standard for nucleic acid amplification test of HP-PRRSV.
作者
原霖
李尔华
董浩
陈亚娜
鞠厚斌
张红丽
王林
宋晓晖
杨林
王传彬
YUAN Lin;LI Er-hua;DONG Hao;CHEN Ya-na;JU Hou-bin;ZHANG Hong-li;WANG Lin;SONG Xiao-hui;YANG Lin;WANG Chuan-bin(China Animal Disease Control Center,Beijing 102600,China;Guangzhou BDS Biological Technology Co.,Ltd,Guangzhou 510000,China;Shanghai Animal Disease Control Centre,Shanghai 201103,China;Zhejiang Animal Disease Control Centre,Hangzhou 310000,China;Beijing Animal Disease Control Centre,Beijing 102600,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2019年第5期826-829,共4页
Chinese Journal of Veterinary Science
基金
动物及动物产品质量安全检测标准物质研制资助项目(2013FY113300-6)
