猪圆环病毒2型质粒标准物质研制OA
Preparation of Reference Material for Genomic Plasmid of Porcine Circovirus Type 2
针对目前我国尚无猪圆环病毒2型(Porcine circovirus type 2,PCV2)核酸标准物质的现状和实际需求,本研究选取08TJ株PCV2为原料,通过分子生物学等技术,研制了PCV2(08TJ株)质粒标准物质.利用所建立的数字PCR方法对其进行均匀性检验和稳定性评估,并委托8家实验室进行合作定值和3家单位试用.结果表明,所研制的标准物质组内和组间均无统计学差异,样品均匀性良好;在-20℃、4℃、25℃和37℃条件下可分别稳定保存12 mo、120 d、14 d和9 d;反复冻融10次后仍稳定;该标准物质标准值±扩增不确定度为(1.1±0.2)×104 copies/µL,与实际样品互通性良好.综上所述,本研究所研制的标准物质均匀性和稳定性良好,定值准确,对提升我国PCV2实验室检测技术水平等具有重要现实意义.
The reference material(RM)for Porcine circovirus type 2(PCV2)is not available currently in China.In this study,RM containing genomic plasmids of PCV2 strain 08TJ was prepared using molecular biology technology.A digital PCR method was established and used to evaluate the homogeneity,stability and characteristics of the RM.Meanwhile,trials of the RM were performed for applications.The homogeneity evaluation showed that there was no statistical difference in within batch and between batch,indicating that the RM we prepared was homogeneous.The monitoring of stability revealed that the RM was stable when it was stored at different temperatures for different time periods,including-20℃for 12 months,4℃for 120 days,25℃for 14 days,and at 37℃for 9 days.Moreover,the RM was stable after it was treated for 10 freezing-thawing cycles.The reference value of the RM was(1.1±0.2)×104 copies/µL.The result of tria l experiments demonstrated that the RM had good commutability with field samples.In conclusion,the RM we prepared had accurate value,good homogeneity and stability.The RM described in this study enhanced the capability of PCV2 detection in veterinary laboratories in China.
张媛;周智;倪建强;孙雨;李琦;翟新验;刘玉良;王传彬;顾小雪;毕一鸣;徐琦;原霖;高旭年;李尔华;钟凤然
中国动物疫病预防控制中心,北京 102618中国动物疫病预防控制中心,北京 102618中国动物疫病预防控制中心,北京 102618中国动物疫病预防控制中心,北京 102618中国动物疫病预防控制中心,北京 102618中国动物疫病预防控制中心,北京 102618中国动物疫病预防控制中心,北京 102618中国动物疫病预防控制中心,北京 102618中国动物疫病预防控制中心,北京 102618中国动物疫病预防控制中心,北京 102618中国动物疫病预防控制中心,北京 102618普道(北京)标准技术有限公司,北京 102629广州邦德盛生物科技有限公司,广州 510700广州邦德盛生物科技有限公司,广州 510700广州邦德盛生物科技有限公司,广州 510700
农业科技
猪圆环病毒标准物质均匀性稳定性
Porcine circovirusreference materialhomogeneitystability
《中国动物传染病学报》 2026 (2)
108-114,7
科技基础性工作专项(2013YF113300)
评论