食品中金黄色葡萄球菌多重PCR检测方法的研究

doi:10.3969/j.issn.1673-1689.2011.01.015

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食品中金黄色葡萄球菌多重PCR检测方法的研究
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                        10.3969/j.issn.1673-1689.2011.01.015
                    
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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(2007B030601003)

Studies on Detection of Staphylococus aureus in Foods by Multiplex PCR

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以耐热核酸酶基因nuc、纤维蛋白原结合蛋白基因clfA和SmaI限制性酶切片段特异序列为靶基因,设计筛选引物,建立并优化了检测金黄色葡萄球菌(Staphylococus aureus,SA)的多重PCR体系。采用10株SA和46株非SA验证了该多重PCR具有很好的特异性。PCR检测的灵敏度在DNA水平上达到1.197 3 ng。人工污染样品,当起始污染量为1.2个/g时,37℃增菌培养6 h即可检出。一共检测了43份样品,检出3份阳性样品,其中有2份阳性样品与传统方法一致,另外一份阳性样品用测序验证。作者建立的多重PCR方法可特异、快速地实现对SA的检测。

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In this manuscript,a multiplex PCR assay for detection of S.aureus was developed by designing three sets of primers that specifically amplify segments of the nuc、clfA and SmaI genes.Analysis of ten S.aureus strains and forty-six non-S.aureus strains demonstrated that this PCR system was specific.The detection limit of the PCR was 1.1973 ng with genomic DNA and 1.2 cfu/g in artificially contaminated bread after enrichment at 37 ℃ for 6 h.Forty-three foods were detected to confirm the accuracy of this method.Among of them,three were found to be positive,and two were consistent with that of the international standard method,and one was through verification by sequenced.The result proven that the specific multiplex PCR assay can be used for detection of S.aureus in foods.

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徐晓可,吴清平,张菊梅,周艳红,杨小鹃.食品中金黄色葡萄球菌多重PCR检测方法的研究[J].食品与生物技术学报,2011,30(1):84-89.

XU Xiao-ke, WU Qing-ping, ZHANG Ju-mei, ZHOU Yan-hong, YANG Xiao-juan. Studies on Detection of Staphylococus aureus in Foods by Multiplex PCR[J]. Journal of Food Science and Biotechnology,2011,30(1):84-89.

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在线发布日期: 2014-06-17
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