[1]李原野,陈世界,林华,等.猪圆环病毒3型微滴式数字PCR检测方法的建立与应用[J].江苏农业学报,2021,(02):389-396.[doi:doi:10.3969/j.issn.1000-4440.2021.02.015]
　LI Yuan-ye,CHEN Shi-jie,LIN Hua,et al.Establishment and application of microdrop digital PCR detection method for porcine circovirus type 3[J].,2021,(02):389-396.[doi:doi:10.3969/j.issn.1000-4440.2021.02.015]
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猪圆环病毒3型微滴式数字PCR检测方法的建立与应用()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2021年02期
页码:
389-396
栏目:
畜牧兽医·水产养殖
出版日期:
2021-04-30

文章信息/Info

Title:
Establishment and application of microdrop digital PCR detection method for porcine circovirus type 3
作者:
李原野1; 陈世界2; 林华2; 张婧2; 安薇2; 陈瑛琪2; 张继宗2; 朱玲1; 3
(1.四川农业大学动物医学院，四川成都611130；2.成都海关检验检疫技术中心，四川成都610041；3.四川农业大学动物疫病与人类健康四川省重点实验室，四川成都611130）
Author(s):
LI Yuan-ye1; CHEN Shi-jie2; LIN Hua2; ZHANG Jing2; AN Wei2; CHEN Ying-qi2; ZHANG Ji-zong2; ZHU Ling1; 3
(1.College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;2.Chengdu Customs Inspection and Quarantine Technology Center, Chengdu 610041, China;3.Sichuan Key Laboratory of Animal Diseases and Human Health, Sichuan Agricultural University, Chengdu 611130, China)
关键词:
猪; 圆环病毒3型; 微滴式数字PCR
Keywords:
pig; circovirus type 3; droplet digital PCR
分类号:
S852.65+9.2
DOI:
doi:10.3969/j.issn.1000-4440.2021.02.015
文献标志码:
A
摘要:
为建立能灵敏、高效、准确诊断猪圆环病毒3型（PCV3）的检测方法，并运用该方法研究PCV3传播特性及组织嗜性，根据GenBank中PCV3 ORF2基因（MF631813.1）的核苷酸序列，设计合成PCV3的引物和探针，通过退火温度以及引物、探针用量和特异性等优化，建立了PCV3微滴式数字PCR检测方法。试验结果表明建立的猪圆环病毒3型微滴式数字PCR检测方法具有较好高的灵敏性、重复性和特异性，最低能检测到1 μl 16.35拷贝的质粒标准品，可用于猪圆环病毒3型的早期诊断。
Abstract:
To establish a detection method that can diagnose porcine circovirus type 3 (PCV3) sensitively, efficiently and accurately and can be used to explore the transmission characteristics and tissue tropism of PCV3, the primers and probes of PCV3 were designed and synthesized according to the nucleotide sequence of PCV3 ORF2 gene (MF631813.1) in GenBank and the annealing temperature, primer and probe dosages and specificity of droplet digital PCR were optimized to establish droplet digital PCR detection method for PCV3. The results showed that, the established droplet digital PCR detection method for porcine circovirus type 3 with high sensitivity, reproducibility and specificity could detect a minimum of 16.35 copies/μl of plasmid standards. The method established in this study can be used for the early diagnosis of the porcine circovirus type 3.

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备注/Memo

备注/Memo:
收稿日期：2020-06-04基金项目：国家“十三五”重点研发计划项目（2018YFD0501102）；四川省重点研发项目（2018NZ0150）作者简介：李原野（1993-），女，四川汉源人，硕士研究生，主要从事动物传染病病原分子生物学研究。(E-mail)544512064@qq.com通讯作者：朱玲，（E-mail）abtczl72@126.com
更新日期/Last Update: 2021-05-10