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目的建立阪崎肠杆菌和沙门菌的多重PCR检测方法。方法针对阪崎肠杆菌外膜蛋白A(ompA)基因和沙门菌属侵袭性抗原保守基因(invA)基因设计引物,建立其多重PCR检测方法,并对反应条件进行优化。结果两对引物分别能扩增出469bp和284bp的目的条带,结果具有高度特异性,反应条件优化后,两菌可同时检出的浓度限度为106cfu/ml。结论该方法敏感、特异,为快速检测阪崎肠杆菌、沙门菌提供了新的有效手段。
Objective To establish a multiplex PCR assay for Enterobacter sakazakii and Salmonella. Methods Primers were designed for Enterobacter sakazakii outer membrane protein A (ompA) gene and Salmonella invasive gene (invA) gene, and their multiplex PCR methods were established. The reaction conditions were optimized. Results Two pairs of primers could amplify the target bands of 469bp and 284bp, respectively. The results were highly specific. After optimizing the reaction conditions, the concentration limit of both bacteria could be detected at 106cfu / ml simultaneously. Conclusion The method is sensitive and specific, which provides a new and effective method for the rapid detection of Enterobacter sakazakii and Salmonella spp.