Novel Development of a qPCR Assay Based on the rpoB Gene for Rapid Detection of Cronobacter spp.

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• 作者：Yuanhong Li ; Qiming Chen ; Hua Jiang ; Yang Jiao ; Fengxia Lu…
• 刊名：Current Microbiology
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：72
• 期：4
• 页码：436-443
• 全文大小：410 KB
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• 作者单位：Yuanhong Li (1) (2)
  Qiming Chen (1)
  Hua Jiang (1) (2)
  Yang Jiao (2)
  Fengxia Lu (1)
  Xiaomei Bie (1)
  Zhaoxin Lu (1)
  
  1. College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang, Jiangsu, 210095, Nanjing, People’s Republic of China
  2. School of Public Health, Xuzhou Medical College, 209 Tongshan Road, Xuzhou, 221004, Jiangsu, People’s Republic of China
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  
• 出版者：Springer New York
• ISSN：1432-0991

文摘

A novel real-time PCR (qPCR) assay with internal amplification control based on the rpoB gene was developed for the detection and quantification of Cronobacter spp. Inclusivity and exclusivity of the qPCR assay were tested on a strain collection containing 19 Cronobacter and 26 non-Cronobacter strains. All Cronobacter strains were successfully identified, whereas no cross-reactivity was observed with non-Cronobacter strains. The sensitivity of the qPCR assay for pure culture and powdered infant formula (PIF) without enrichment was 3.44 log CFU/ml(g) (2.74 × 103 CFU/ml(g)). When the qPCR assay was applied to artificially contaminated PIF after a 12-h enrichment step, as few as 0.03 log CFU/ml (1.06 × 100 CFU/ml) of C. sakazakii could be detected. The limit of detection of the qPCR assay was not reduced by the presence of 8 log CFU/ml of Salmonella Enteritidis in PIF. A total of 70 food samples were analyzed for the presence of Cronobacter spp., out of which 3 dry cereal products, 5 maternal milk, and 1 infant food formula were found as positive by qPCR. The results obtained by qPCR were consistent with those obtained by culture-based method. Results from this study demonstrate that the qPCR assay is a rapid, specific, and accurate method suitable for Cronobacter detection in foods.