人博卡病毒微滴数字PCR定量方法的建立  

Establishment of Droplet Digital Polymerase Chain Reaction for Quantifying Human Bocavirus

作　　者：邓幼平[1] 刘颖娟[2] 王笑臣 刘佳明 杨占秋[3] 赵东赤[1] Deng Youping;Liu Yingjuan;Wang Xiaochen;Liu Jiaming;Yang Zhanqiu;Zhao Dongchi(Department of Pediatrics,Zhongnan Hospital of Wuhan University,Wuhan 430071,China;Clinical Laboratory,Zhongnan Hospital of Wuhan University,Wuhan 430071,China;State Key Laboratory of Virology,Institute of Medical Virology,School of Medicine,Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院儿科,武汉430071 [2]武汉大学中南医院检验科,武汉430071 [3]武汉大学医学院医学病毒学研究所,病毒学国家重点实验室,武汉430071

出　　处：《中南民族大学学报（自然科学版）》2018年第3期32-36,共5页Journal of South-Central University for Nationalities：Natural Science Edition

基　　金：国家自然科学基金资助项目(81101306)

摘　　要：为准确、快速地从临床鼻咽拭子样本中筛选出人博卡病毒(HBoV)感染的阳性样本,以HBoV-sh9基因的保守区序列为检测靶标合成引物和探针,建立了微滴数字PCR(dd PCR)检测HBoV的方法.结果表明:HBoV dd PCR方法具有较高的特异性,在(0.5~6.8)×10~3copies/μL HBoV DNA浓度范围内,具有良好的线性和精确度,定量限低至0.5 copies/μL.182份临床样本检测结果显示:HBoV感染的阳性率为8.24%.此建立的HBoV dd PCR方法在定量限、准确性和重复性上较常规PCR优势明显,不受样品基质和扩增效率的影响.In order to accurately and quickly detect positive specimens of human bocavirus(HBoV)infection from clinical nasopharyngeal swab samples,primers and probes were synthesized by using the conserved region sequence of HBoV-sh9 gene as the target and the method of droplet digital polymerase chain reaction(ddPCR)for HBoV detection was established.The results showed that ddPCR method had high specificity,good linearity and precision within the HBoV DNA range of(0.5~6.8)×10 3 copies/μL,and the detection limit was as low as 0.5 copies/μL.182 clinical samples were tested and a positive HBoV infection rate of 8.24%was found.This HBoV ddPCR method had obvious advantages in quantitative limit,accuracy and repeatability compared with the conventional PCR and was not affected by sample matrix and amplification efficiency.

关 键 词：微滴数字聚合酶链式反应 人博卡病毒 拷贝数 阳性率 

分 类 号：O655.2[理学—分析化学]