Genome Mining of Novel Targets and Construction of Ladder-shaped melting temperature isothermal amplification Assays for the Identification of Cronobacter sakazakii and Cronobacter malonaticus.
Da Ma, Ling Deng, Liangpi Huang, Aoxiang Peng, Wanglai Bi, Rui Li
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引用次数: 0
Abstract
Cronobacter species are potential pathogens that can contaminate powdered infant formula. C. sakazakii and C. malonaticus are the most common species of Cronobacter associated with infections. This study mined new molecular targets for the detection of C. sakazakii and C. malonaticus by using comparative genome approaches. Specific target genes mngB and ompR were obtained and used to detect C. sakazakii and C. malonaticus, respectively. A novel detection method, termed ladder-shape melting temperature isothermal amplification (LMTIA), was developed and evaluated. The detection limit for pure C. sakazakii DNA was 1 pg per reaction and 1 fg per reaction for C. malonaticus. The C. sakazakii, C. malonaticus, and the reference stains were all correctly identified. The amplicons can be successfully visualized and identified by naked eyes when hydroxy naphthol blue dye (HNB dye) was used in the reaction. Therefore, the LMTIA assays developed in this study showed potential application for microorganism identification and detection.
期刊介绍:
Foodborne Pathogens and Disease is one of the most inclusive scientific publications on the many disciplines that contribute to food safety. Spanning an array of issues from "farm-to-fork," the Journal bridges the gap between science and policy to reduce the burden of foodborne illness worldwide.
Foodborne Pathogens and Disease coverage includes:
Agroterrorism
Safety of organically grown and genetically modified foods
Emerging pathogens
Emergence of drug resistance
Methods and technology for rapid and accurate detection
Strategies to destroy or control foodborne pathogens
Novel strategies for the prevention and control of plant and animal diseases that impact food safety
Biosecurity issues and the implications of new regulatory guidelines
Impact of changing lifestyles and consumer demands on food safety.