Rapid detection of Enterobacter sakazakii using TaqMan real-time PCR assay.
J Microbiol Biotechnol
; 17(3): 516-9, 2007 Mar.
Article
em En
| MEDLINE
| ID: mdl-18050957
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.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
RNA Ribossômico 16S
/
Reação em Cadeia da Polimerase
/
Técnicas de Tipagem Bacteriana
/
Taq Polimerase
/
Cronobacter sakazakii
/
Alimentos Infantis
Tipo de estudo:
Diagnostic_studies
/
Evaluation_studies
/
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
J Microbiol Biotechnol
Ano de publicação:
2007
Tipo de documento:
Article