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首页> 外文期刊>Journal of Clinical Microbiology >Simultaneous Analysis of Multiple Staphylococcal Enterotoxin Genes by an Oligonucleotide Microarray Assay
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Simultaneous Analysis of Multiple Staphylococcal Enterotoxin Genes by an Oligonucleotide Microarray Assay

机译：通过寡核苷酸芯片技术同时分析多个葡萄球菌肠毒素基因

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Staphylococcal enterotoxins (SEs) are a family of 17 major serological types of heat-stable enterotoxins that are one of the leading causes of gastroenteritis resulting from consumption of contaminated food. SEs are considered potential bioweapons. Many Staphylococcus aureus isolates contain multiple SEs. Because of the large number of SEs, serological typing and PCR typing are laborious and time-consuming. Furthermore, serological typing may not always be practical because of antigenic similarities among enterotoxins. We report on a microarray-based one-tube assay for the simultaneous detection and identification (genetic typing) of multiple enterotoxin (ent) genes. The proposed typing method is based on PCR amplification of the target region of the ent genes with degenerate primers, followed by characterization of the PCR products by microchip hybridization with oligonucleotide probes specific for each ent gene. We verified the performance of this method by using several other techniques, including PCR amplification with gene-specific primers, followed by gel electrophoresis or microarray hybridization, and sequencing of the enterotoxin genes. The assay was evaluated by analysis of previously characterized staphylococcal isolates containing 16 ent genes. The microarray assay revealed that some of these isolates contained additional previously undetected ent genes. The use of degenerate primers allows the simultaneous amplification and identification of as many as nine different ent genes in one S. aureus strain. The results of this study demonstrate the usefulness of the oligonucleotide microarray assay for the analysis of multitoxigenic strains, which are common among S. aureus strains, and for the analysis of microbial pathogens in general.
机译：葡萄球菌肠毒素（SEs）是热稳定肠毒素的17种主要血清学类型的家族，是食用受污染食物导致肠胃炎的主要原因之一。 SE被认为是潜在的生物武器。许多金黄色葡萄球菌分离株含有多个SE。由于SE数量众多，血清学分型和PCR分型既费力又费时。此外，由于肠毒素之间的抗原相似性，血清学分型可能并不总是可行的。我们报告了基于微阵列的单管测定法，用于同时检测和鉴定多种肠毒素（ ent ）基因。提出的分型方法是基于用简并引物PCR扩增 ent 基因的目标区域，然后通过与每种 ent特异的寡核苷酸探针进行微芯片杂交来表征PCR产物基因。我们通过使用其他几种技术验证了该方法的性能，包括使用基因特异性引物进行PCR扩增，然后进行凝胶电泳或微阵列杂交，并对肠毒素基因进行测序。通过分析先前鉴定的含有16个 ent 基因的葡萄球菌分离株来评估该测定。芯片检测表明，其中一些分离株还含有其他先前未检测到的 ent 基因。简并引物的使用允许在一个 S中同时扩增和鉴定多达9个不同的 ent 基因。金黄色葡萄球菌这项研究的结果证明了寡核苷酸微阵列测定法可用于分析多毒素菌株，这在 S中很常见。金黄色葡萄球菌菌株，一般用于分析微生物病原体。

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