A precise 5' nuclease (TaqMan) real-time PCR was developed and validated in house for the specific detection of Enterobacter sakazakii isolates. Specifically designed nonpatented primers and a hydrolysis (TaqMan) probe were used to target the 16S rRNA gene. All 27 E. sakazakii and 141 non-E. sakazakii strains tested with the real-time PCR were identified correctly. To monitor false-negative results, an internal amplification control was coamplified with the same primers used for the E. sakazakii DNA. The detection probability of the assay was 56% when an E. sakazakii cell suspension containing 10(2) CFU/ml was used as template in the PCR (0.5 CFU per reaction) and 100% with a 10(3) CFU/ml suspension. This PCR assay should be very useful for the diagnostic detection of E. sakazakii in foods, especially powdered infant formula, after cultural enrichment.
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页码:1623 / 1627
页数:5
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[1]
Abu Al-Soud W, 1998, APPL ENVIRON MICROB, V64, P3748
机构:
Univ Massachusetts, Sch Med, Div Infect Dis & Immunol, Dept Med,UMass Mem Med Ctr, Worcester, MA 01655 USAUniv Massachusetts, Sch Med, Div Infect Dis & Immunol, Dept Med,UMass Mem Med Ctr, Worcester, MA 01655 USA
机构:
Univ Massachusetts, Sch Med, Div Infect Dis & Immunol, Dept Med,UMass Mem Med Ctr, Worcester, MA 01655 USAUniv Massachusetts, Sch Med, Div Infect Dis & Immunol, Dept Med,UMass Mem Med Ctr, Worcester, MA 01655 USA