NDLI: Development of multiplex real-time PCR with Internal amplification control for simultaneous detection of Salmonella and Cronobacter in powdered infant formula.

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Development of multiplex real-time PCR with Internal amplification control for simultaneous detection of Salmonella and Cronobacter in powdered infant formula.

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Development of multiplex real-time PCR with Internal amplification control for simultaneous detection of Salmonella and Cronobacter in powdered infant formula.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Hyeon, Ji-Yeon Park, Chankyu Choi, In-Soo Holt, Peter S. Seo, Kun-Ho
Description	Country affiliation: South Korea Author Affiliation: Hyeon JY ( Department of Public Health, College of Veterinary Medicine, Konkuk University, Seoul 143-701, South Korea.)
Abstract	Contamination of powdered infant formula (PIF) by the bacteria Cronobacter spp. and Salmonella enterica was deemed a matter of great concern by the World Health Organization and the Food and Agriculture Organization of the United Nations in 2004. Therefore, we developed a rapid and sensitive multiplex real-time PCR assay for the simultaneous detection of Cronobacter and Salmonella in PIF. In addition, an internal amplification control (IAC) was also included for exclusion of false negative results in this study. The quantitative detection range for pure cultures in this optimized multiplex real-time PCR assay was 10³ to 108 CFU/ml for both Salmonella and Cronobacter. When our established multiplex real-time PCR system was applied to artificially contaminated PIF, the detection limit was 10³ CFU/ml for Salmonella and Cronobacter without enrichment. The commercial PIF was then inoculated with Salmonella and Cronobacter at 10, 1 and 0.1 CFU per gram of formula and the single enrichment broth samples were analyzed by multiplex real-time PCR after enrichment for 9, 12, and 24 h. At 12 h post-enrichment, we could detect Salmonella and Cronobacter at initial inoculation levels of approximately 0.1 CFU/g in PIF. Additionally, stable fluorescent IAC signals could be assessed between 29 and 34 cycles of PCR amplification. Results from this study showed that the multiplex real-time PCR assay is an effective method for the rapid and simultaneous detection and quantification of Cronobacter and Salmonella in PIF.
File Format	HTM / HTML
ISSN	01681605
Issue Number	1
Volume Number	144
e-ISSN	18793460
Journal	International Journal of Food Microbiology
Language	English
Publisher	Elsevier
Publisher Date	2010-11-15
Publisher Place	Netherlands
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Salmonella Research Support, Non-u.s. Gov't Discipline Microbiology Sensitivity And Specificity Infant Formula Humans Isolation & Purification Infant Food Microbiology Journal Article Time Factors Polymerase Chain Reaction Genetics Discipline Nutritional Sciences Methods Enterobacteriaceae
Content Type	Text
Resource Type	Article
Subject	Medicine Microbiology Safety, Risk, Reliability and Quality Food Science

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Development of multiplex real-time PCR with Internal amplification control for simultaneous detection of Salmonella and Cronobacter in powdered infant formula.

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