鸡白痢沙门氏菌直接-多重PCR检测体系的建立
Establishment of a direct multiplex PCR detection system of Salmonella pullorum
作者：程玲玲(四川出入境检验检疫局检验检疫技术中心)；严伟(四川大学生命科学学院 动物疫病防控与食品安全四川省重点实验室)；侯若彤(四川大学生命科学学院 动物疫病防控与食品安全四川省重点实验室)；帅培强(四川出入境检验检疫局检验检疫技术中心)；张彪(成都市食品药品检验研究院)；白林含(四川大学生命科学学院 动物疫病防控与食品安全四川省重点实验室)
Author：CHEN Ling-Ling(lnspection and Quarantine Technical Center of Sichuan Entry-exit lnspection and Quarantine Bureau)；YAN Wei(Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University)；HOU Ruo-Tong(Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University)；SHUAI Pei-Qiang(lnspection and Quarantine Technical Center of Sichuan Entry-exit lnspection and Quarantine Bureau)；ZHANG Biao(Chengdu Institutes for Food and Drug Control)；BAI Lin-Han(Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University)
收稿日期：2017-09-29 年卷（期）页码：2019,56(1):149-154
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：鸡白痢沙门氏菌；毒力质粒 pSPUV；特异检测引物；DNA聚合酶共用反应缓冲液；直接三重PCR检测体系
Key words：Salmonella pullorum; Virulence plasmid pSPUV; Specific detection primers, Common reaction buffer of DNA polymerase; Direct triple PCR system
基金项目：国家蛋鸡产业技术体系岗位科学家项目（CARS-41-K09）
中文摘要
通过对鸡白痢沙门氏菌毒力质粒pSPUV上入侵质粒抗原J蛋白和质粒共转移调控因子的编码基因 ipaJ 和 traJ 的序列分析,在特异区域设计PCR引物,经沙门氏菌属不同种及常见肠道致病菌的交叉验证,筛选出三对鸡白痢沙门氏菌特异检测引物ipaJ-417、traJ-387、traJ-476. 并且调试出一种利用两种DNA聚合酶的直接三重PCR检测体系:invA-211/traJ-387/16S-495,可以对鸡白痢沙门氏菌进行快速准确的鉴定.
英文摘要
In this paper, we screened out three pairs of specific detection primers of Salmonella pullorum, ipaJ 417, traJ-387, and traJ-476, by the blast analysis of the coding genes ipaJ and traJ which coded the invasion plasmid antigen protein J and plasmid co-transfer regulatory factor in Salmonella pullorum virulence plasmid Pspuv. And these PCR primers were designed in special regions and identificated in the different species of Salmonella and common intestinal pathogens. And a common reaction buffer of two kinds of DNA polymerase for the direct triple PCR system was also established (invA-211/traJ-387/16S-495). This direct triple PCR system can be used for fast and accurate identification of Salmonella pullorum.
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