| 食品中志贺菌特异性二重PCR检测方法的研究 |
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| 引用本文： | 徐晓可,吴清平,张菊梅,周艳红,杨小鹃. 食品中志贺菌特异性二重PCR检测方法的研究[J]. 中国卫生检验杂志, 2010, 0(11) |
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| 作者姓名： | 徐晓可 吴清平 张菊梅 周艳红 杨小鹃 |
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| 作者单位： | 广东省微生物研究所广东省菌种保藏与应用重点实验室广东省微生物应用新技术公共实验室; |
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| 基金项目： | 科技型中小企业技术创新基金(09C26214402098); 广东省计划项目(2008A040101001) |
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| 摘 要： | 目的:建立二重PCR方法快速检测食品中的志贺菌(Shigella spp.)。方法:以侵袭性质粒抗原H基因ipaH和铁载体基因iuc为靶基因,筛选引物,建立二重PCR体系。对3株志贺菌和28株非志贺菌进行特异性检测。梯度稀释志贺菌基因组DNA,以不同稀释度DNA作PCR扩增。在孜然牛肉中以不同菌量人工污染,不同增菌时间培养,提取DNA进行PCR扩增。应用该方法检测实际样品。结果:以ipaH、iuc为靶基因的2对引物对志贺菌的检出有很好的特异性。PCR检测的灵敏度在DNA水平上达到78.73 pg。人工污染样品,当起始污染量为0.56 cfu/g时,37℃增菌培养10 h即可检出。一共检测了18份样品,未检出志贺菌,与传统方法一致。利用该方法检测了一份盲样冻干样品,检出志贺菌,与实际结果相符。结论:建立了适用于食品中志贺菌检测的特异灵敏二重PCR方法。
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| 关 键 词： | 志贺菌 二重PCR 食品 |
Studies on specific detection of Shigella spp. in foods by duplex PCR |
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| XU Xiao-ke,WU Qing-ping,ZHANG Ju-mei,ZHOU Yan-hong,YANG Xiao-juan. Studies on specific detection of Shigella spp. in foods by duplex PCR[J]. Chinses Journal of Health Laboratory Technology, 2010, 0(11) |
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| Authors: | XU Xiao-ke WU Qing-ping ZHANG Ju-mei ZHOU Yan-hong YANG Xiao-juan |
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| Affiliation: | XU Xiao-ke,WU Qing-ping,ZHANG Ju-mei,ZHOU Yan-hong,YANG Xiao-juan(Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application,Guangdong Provincial Public Laboratory of Microbial Application of New Technologies,Guangdong Institute of Microbiology,Guangzhou 510070,China) |
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| Abstract: | Objective:A rapid dulplex PCR assay was developed to detect Shigella spp.in foods.Methods: A dulplex PCR assay for detection of Shigella spp.was developed by designing two sets of primers that specifically amplify segments of the ipaH and iuc genes.Three Shigella spp.(Shigella dysenteriae,Shigella flexneri,Shigella sonnei) strains and tweenty-eight non-Shigella spp.strains were amplified by PCR to confirm the specificity.The Shigella sonnei genomic DNA was serially diluted and subjected to PCR amplification... |
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| Keywords: | Shigella spp. Duplex PCR Foods |
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