Detection of Enterobacter sakazakii strains by real-time PCR

被引:28
作者
Malorny, B
Wagner, M
机构
[1] Fed Inst Risk Assessment, D-12277 Berlin, Germany
[2] Inst Milk Hyg Milk Technol & Food Sci, A-1210 Vienna, Austria
关键词
D O I
10.4315/0362-028X-68.8.1623
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:1623 / 1627
页数:5
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