Design of a 5′ exonuclease-based real-time PCR assay for simultaneous detection of Bacillus licheniformis, members of the 'B-cereus group' and B-fumarioli in gelatine

被引:15
作者
De Clerck, E
Van Mol, K
Jannes, G
Rossau, R
De Vos, P
机构
[1] Univ Ghent, Dept Biochem Physiol & Microbiol, Microbiol Lab, B-9000 Ghent, Belgium
[2] Innogenet NV, Ghent, Belgium
关键词
gelatinase; identification; public health; SmartCycler; Taqman;
D O I
10.1111/j.1472-765X.2004.01550.x
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
Aims: The design of a fast, sensitive and specific detection method for Bacillus licheniformis, members of the 'B. cereus group' and B. fumarioli in gelatine. Methods and Results: Specific Taqman probes were designed and tested in a real-time PCR setting. A specific fluorescent signal could be obtained for all gelatine isolates attributed to these species in one single real-time PCR reaction. After sample preparation, a gelatine sample spiked with 1 CFU provided enough template DNA for a significant signal. Conclusion: The potential of a real-time PCR assay for simultaneous detection of B. licheniformis, members of the 'B. cereus group' and B. fumarioli in gelatine is demonstrated. Significance and Impact of the Study: Implementation of the assay in gelatine producing plants may shorten delivery terms and inform on hazards to public health and suitable remediation procedures.
引用
收藏
页码:109 / 115
页数:7
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