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[用于同时鉴定金黄色葡萄球菌和检测食物中毒暴发分离株中葡萄球菌肠毒素的多重PCR策略]

[Multiplex PCR strategy for the simultaneous identification of Staphylococcus aureus and detection of staphylococcal enterotoxins in isolates from food poisoning outbreaks].

作者信息

Brizzio Aníbal A, Tedeschi Fabián A, Zalazar Fabián E

机构信息

Servicio de Laboratorio, Agencia Santafesina de Seguridad Alimentaria (ASSAL), Santa Fe, Argentina.

出版信息

Biomedica. 2013 Jan-Mar;33(1):122-7. doi: 10.1590/S0120-41572013000100015.

Abstract

INTRODUCTION

Staphylococcal food poisoning is the most frequent type of food poisoning around the world. Staphylococcus aureus enterotoxins cause significant loss of water in the intestinal lumen, followed by vomiting and diarrhea.

OBJECTIVE

To report a fast, reliable and inexpensive strategy based on multiplex PCR for the simultaneous identification of S. aureus and detection of five classical S. aureus enterotoxin genes ( sea, seb, sec, sed, see ) in Staphylococcus spp. strains isolated from food poisoning outbreaks.

MATERIALS AND METHODS

We analyzed isolates from 12 food poisoning outbreaks occurred in Santa Fe province (Argentina). Isolation and phenotypic characterization were carried out by standard procedures. Genotypic analysis was performed by multiplex PCR, using primers for nuc , sea-see and 16S rRNA genes simultaneously.

RESULTS

Of all the strains tested, 58% were found to carry toxigenic genes. Sea and seb toxins were found at the same percentage (29%) while sec, sed and see genes were found in a lower and identical proportion (14%). We did not find more than one different type of S. aureus enterotoxin in the isolates analyzed.

CONCLUSIONS

The multiplex PCR strategy designed in this work has enabled us to identify strains of S. aureus and detect -at the same time- their enterotoxigenic ability. At present, our efforts are devoted to the detection of genes encoding enterotoxins other than the classical ones, in order to know their impact on staphylococcal food poisoning, as well as to investigate their relevance to our country's public health.

摘要

引言

葡萄球菌食物中毒是全球最常见的食物中毒类型。金黄色葡萄球菌肠毒素会导致肠腔内大量失水,继而引发呕吐和腹泻。

目的

报告一种基于多重聚合酶链反应(multiplex PCR)的快速、可靠且经济的策略,用于同时鉴定从食物中毒暴发中分离出的葡萄球菌属菌株中的金黄色葡萄球菌,并检测五个经典的金黄色葡萄球菌肠毒素基因(sea、seb、sec、sed、see)。

材料与方法

我们分析了来自阿根廷圣菲省发生的12起食物中毒暴发的分离株。通过标准程序进行分离和表型特征鉴定。使用针对nuc、sea-see和16S rRNA基因的引物同时进行多重聚合酶链反应(multiplex PCR)以进行基因分型分析。

结果

在所有测试菌株中,58%被发现携带产毒基因。发现Sea和Seb毒素的比例相同(29%),而Sec、Sed和See基因的比例较低且相同(14%)。在分析的分离株中未发现超过一种不同类型的金黄色葡萄球菌肠毒素。

结论

本研究设计的多重聚合酶链反应(multiplex PCR)策略使我们能够鉴定金黄色葡萄球菌菌株,并同时检测其产肠毒素能力。目前,我们致力于检测除经典肠毒素基因以外的其他编码肠毒素的基因,以了解它们对葡萄球菌食物中毒的影响,以及调查它们与我国公共卫生的相关性。

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