在线阅读 --自然科学版 2014年5期《乳粉中常见致病菌的多重荧光PCR检测》
乳粉中常见致病菌的多重荧光PCR检测--[在线阅读]
秦丽1, 周巍1,2, 张明3, 杨帆1, 贾昭斌1, 张岩1
1. 河北省食品质量监督检验研究院, 河北 石家庄 050051;
2. 河北农业大学 食品科技学院, 河北 保定 071000;
3. 沈阳辉山国家农业科技园区 管理委员会, 辽宁 沈阳 110164
起止页码: 512--517页
DOI: 10.11826/j.issn.1000-5854.2014.05.015
摘要
利用多重荧光PCR技术快速检测乳粉中的沙门氏菌、金黄色葡萄球菌、志贺氏菌。以沙门氏菌的invA基因、志贺氏菌的ipaH基因、金黄色葡萄球菌的nuc基因为靶基因,选择特异性引物,以参照菌种为对象,验证引物的特异性,建立多重荧光PCR检测方法,人工添加沙门氏菌、金黄色葡萄球菌、志贺氏菌,确定方法的检出限。结果表明该方法特异性强、灵敏度高,乳粉中检测的灵敏度为1CFU/mL,可在8h内完成对乳品中沙门氏菌、金黄色葡萄球菌、志贺氏菌的检测。

Detection of Common Foodborne Pathogens in Emulsifiable Dust by Multiplex RT-PCR Assay
QIN Li1, ZHOU Wei1,2, ZHANG Ming3, YANG Fan1, JIA Zhaobin1, ZHANG Yan1
1. Hebei Institute of Food Quality Supervision Inspection and Research, Hebei Shijiazhuang 050051, China;
2. College of Food Science and Technology, Agricultural University of Hebei, Hebei Baoding 071000, China;
3. Administrative Committee of Shenyang Huishan National Agricultural Science Park, Liaoning Shenyang 110164, China
Abstract:
A multiplex RT-PCR assay was designed for prompt detection of Staphylococcus aureus,Salmonella spp, and Shigella spp in milk powder.In this study,the target genes were the invA of Salmonella,the ipaH of Shigella and the nuc of Staphylococcus,and specific primers were designed accordingly.Staphylococcus aureus,Salmonella spp,and Shigella spp were artificially added to determine the detection limit of the method.The results indicated that detection of Staphylococcus aureus,Salmonella spp,and Shigella spp in milk powder by multiplex RT-PCR was of strong specificity and high sensitivity,and the sensitivity of the multiplex RT-PCR was 1CFU/mL.The assay was capable of detecting Staphylococcus aureus,Salmonella spp,and Shigella spp in milk powder within 8hours.

收稿日期: 2014-4-9
基金项目: 河北省质量技术监督局科研项目(090106)

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