作 者：
原霖;訾占超;亢文华;李晓霞;汪葆玥;王静;韩焘;陈亚娜;倪建强;翟新验
关键词：
猪圆环病毒Ⅰ型;微滴数字PCR;检测方法
摘 要：
以猪圆环病毒1型ORF1基因为靶基因,建立了可对其准确定量的微滴数字PCR(droplet digital PCR,ddPCR)方法。对ddPCR反应中的探针浓度进行了优化。结果显示:ddPCR反应中的最佳探针浓度为250nmol/L;ddPCR方法线性相关系数(R2)为0.999,呈良好的线性关系;检测限为7.8copies/20μL;变异系数为2.7%;与其他病毒无交叉反应。通过对临床样品的检测,检测结果与实时荧光PCR结果一致。结果表明:新建立的ddPCR方法敏感性高、特异性强,可用于猪圆环病毒1型的定量检测。
译 名：
Development of droplet digital PCR for the detection of porcine circovirus type 1
作 者：
YUAN Lin;ZI Zhan-chao;KANG Wen-hua;LI xiao-xia;WANG Bao-yue;WANG Jing;HAN Tao;CHEN Ya-na;NI Jian-qiang;ZHAI Xin-yan;OIE Reference Laboratory for Porcine Reproductive and Respiratory Syndrome,China Animal Disease Prevention and Control Center;College of Veterinary Medicine,China Agricultural University;
单 位：
OIE Reference Laboratory for Porcine Reproductive and Respiratory Syndrome,China Animal Disease Prevention and Control Center%College of Veterinary Medicine,China Agricultural University
关键词：
porcine circovirus type 1;;droplet digital PCR;;detection
摘 要：
A droplet digital polymerase chain reaction(ddPCR)method,based on ORF1 gene of porcine circovirus type 1,PCV1,was developed to improve the detection and quantification of PCV1.The probe concentration in ddPCR was optimized.The optimized probe concentration was250 nmol/L.The R2 value of ddPCR standard curve was 0.999,with a good linear response.The limit of detection was 7.8 copies/20μL,the coefficient of variation was 2.7%,the ddPCR was specific for detecting PCV1.The sensitivity of ddPCR was equal to real time quantitative PCR and qPCR by detecting the field samples.The results indicate that the new ddPCR assay provides a specific,sensitive tool for quantitative detection of PCV1.
