Rapid detection of Enterobacter sakazakii using TaqMan real-time PCR assay

Eun Sil Kang, Yong Suk Nam, Kwang Won Hong

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)516-519
Number of pages4
JournalJournal of Microbiology and Biotechnology
Volume17
Issue number3
StatePublished - Mar 2007

Keywords

  • 16S rRNA
  • Enterobacter sakazakii
  • Rapid detection
  • Real-time PCR

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