猪繁殖与呼吸综合征病毒美洲经典株国家核酸标准物质的研制  

Development of the national nucleic acid reference material of NA-PRRSV

作　　者：原霖[1] 高旭年 董浩[1] 陈亚娜[1] 闫若潜[3] 兰德松[4] 宋晓晖[1] 吴佳俊[1] 杨林[1] 王传彬[1] YUAN Lin;GAO Xunian;DONG Hao;CHEN Yana;YAN Ruoqian;LAN Desong;SONG Xiaohui;WU Jiajun;YANG Lin;WANG Chuanbin(The China Animal Disease Control Center,Beijing 102600,China;Guangzhou BDS Biological Technology Co.,Ltd,Guangzhou 510000,China;The Henan Animal Disease Control Center,Zhengzhou 450016,China;The Liaoning Animal Disease Control Center,Shenyang 110000,China)

机构地区：[1]中国动物疫病预防控制中心,北京102618 [2]广州邦德盛生物科技有限公司,广东广州510000 [3]河南省动物疫病预防控制中心,河南郑州450016 [4]辽宁省动物疫病预防控制中心,辽宁沈阳110000

出　　处：《畜牧与兽医》2018年第11期78-82,共5页Animal Husbandry & Veterinary Medicine

基　　金：动物及动物产品质量安全检测标准物质研制(2013FY113300-6)

摘　　要：为了研制猪繁殖与呼吸综合征病毒（porcine reproductive and respiratory syndrome virus,PRRSV）美洲经典株国家核酸标准物质,将美洲经典株代表PRRSV VR2332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。制备后进行均匀性评估、稳定性评估、数字PCR法定值和临床试用。结果表明制备的标准物质的定值为（2. 06±0. 31）×104copies/μL;样品均匀;-20℃稳定保存12个月以上、4℃稳定2个月以上。经全国标准物质管理委员会评审,该制备物达到了国家二级标准物质的要求,可以作为核酸扩增检测的PRRSV美洲经典株核酸标准物质。To prepare the national reference material of NA-PRRSV for nucleic acid amplification technology, Marc145 cell lines were in- fected by NA-PRRSV （VR2332）. 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 ma- terials were valued using the digital PCR, and their clinical applicability was evaluated. The results indicated that the value of the reference material was （2.06±0. 31）×104 copies/μL. The prepared materials were stable at -20 ~C for at least 12 months, and at 4 ℃ for more than 2 months. The prepared material of PRRSV passed the review of the National Administrative Committee for CRM＇ s who believed that it reached the requirements of the national secondary reference. This reference material could be used as the standard material for the nucleic acid amplification test on NA-PRRSV.

关 键 词：猪繁殖与呼吸综合征病毒美洲经典株 核酸扩增 标准物质 

分 类 号：S855.3[农业科学—临床兽医学]