应用猪繁殖与呼吸综合征核酸标准物质比较5种核酸提取试剂盒  

Comparison of Five Kinds of Nucleic Acid Extraction Kits Based on Nucleic Acid Reference Materials of PRRSV

作　　者：徐琦[1] 韩焘[1] 原霖[1] 张硕[1] 李晓霞 刘颖昳[1] 毕一鸣[1] 顾小雪[1] Xu Qi;Han Tao;Yuan Lin;Zhang Shuo;Li Xiaoxia;Liu Yingyi;Bi Yiming;Gu Xiaoxue(China Animal Disease Control Center,Beijing 102618,China)

机构地区：[1]中国动物疫病预防控制中心,北京102618

出　　处：《中国动物检疫》2020年第12期120-123,共4页China Animal Health Inspection

基　　金：国家重点研发计划项目(2018YFD0500800)。

摘　　要：为比较不同核酸提取试剂盒对猪繁殖与呼吸综合征病毒(PRRSV)核酸的提取效率,使用国家标准物质“猪繁殖与呼吸综合征病毒美洲经典株核酸标准物质”作为模拟样品,分别使用试剂盒A、B、C(磁珠法)以及试剂盒D、E(柱式法)等5种核酸提取试剂盒进行核酸提取,然后通过荧光RT-PCR和数字RT-PCR方法评估提取效率,并分析不同基质对提取效率的影响。结果显示:5种试剂盒均能有效提取PRRSV RNA,其中磁珠法的提取效率高于柱式法;试剂盒B和C对PRRSV RNA的提取效率较高;基质会对提取效率造成影响,精液和抗凝血对试剂盒提取效率的影响较大。综上,试剂盒B、C均可适用于猪繁殖与呼吸综合征临床样品的核酸提取。实际操作中,应根据实验室条件、样品数量、样品类型和时间要求等因素,选择应用合适的提取试剂盒。In order to compare the efficiency of various nucleic acid extraction kits for RNA of porcine reproductive and respiratory syndrome virus(PRRSV),national reference material“PRRSV American classic strain nucleic acid reference material”was respectively extracted by five kinds of kits(three magnetic bead kits including A,B and C and two column kits including D and E),and their efficiency were evaluated by fluorescent RT-PCR and digital RTPCR.Then,the effect of different substrates on the efficiency was analyzed.The results showed that all the kits could effectively extract RNA of PRRSV,in which,the efficiency of magnetic bead kits was higher than that of column kits;the efficiency of B and C was higher;the efficiency could be affected by substrates,especially semen and anticoagulant.In conclusion,both B and C were appropriate for extraction of nucleic acids from clinical samples of PRRSV.Therefore,appropriate kits should be used subject to laboratory conditions,number of samples,types of samples,urgency of cases and other factors.

关 键 词：猪繁殖与呼吸综合征 核酸提取 标准物质 数字PCR 

分 类 号：S852.65[农业科学—基础兽医学]