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KMID : 0545120070170030516
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Journal of Microbiology and Biotechnology
2007 Volume.17 No. 3 p.516 ~ p.519
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Rapid Detection of Enterobacter sakazakii Using TaqMan Real-Time PCR Assay
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Kang Eun-Sil
Hong Kwang-Won
Nam Yong-Suk
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Abstract
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Enterobacter sakazakii is an emerging food pathogen, which induces severe meningitis and sepsis in neonates and infants, with a high fatality rate. The disease is generally associated with the ingestion of contaminated infant formula. In this study, we describe the development of a real-time PCR protocol to identify E. sakazakii using a TaqMan probe, predicated on the nucleotide sequence data of the 16S rRNA gene obtained from a variety of pathogens. To detect E. sakazakii, four primer sets and one probe were designed. Five strains of E. sakazakii and 28 non-E. sakazakii bacterial strains were used in order to ensure the accuracy of detection. The PCR protocol successfully identified all of the E. sakazakii strains, whereas the 28 non-E. sakazakii strains were not detected by this method. The detection limits of this method for E. sakazakii cells and purified genomic DNA were 2.3 CFU/ assay and 100 fg/assay, respectively. These findings suggest that our newly developed TaqMan real-time PCR method should prove to be a rapid, sensitive, and quantitative method for the detection of E. sakazakii.
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KEYWORD
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Enterobacter sakazakii, real-time PCR, 16S rRNA, rapid detection
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