Journal
FOOD CONTROL
Volume 125, Issue -, Pages -Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.foodcont.2021.107896
Keywords
Cronobacter species; Bioinformatics; Target mining; PCR; Multiplex PCR
Categories
Funding
- National Natural Science Foundation of China [31972175]
- Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program [2017BT01S174]
- Guangdong Academy of Sciences Special Project of Implementing InnovationDriven Development Capacity Building [2019GDSYL0201001]
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This study successfully identified species-specific molecular targets for different Cronobacter species using a bioinformatics approach and developed a multiplex PCR method for species identification. The PCR and mPCR assays showed excellent specificity and sensitivity for detecting Cronobacter species, suggesting their potential for use in food inspection and clinical diagnosis.
Cronobacter species are opportunistic pathogens associated with severe infections in immunocompromised individuals. Species differentiation is essential as the sensitivity of different species to chemical reagents, antibiotics, and virulence factors may be diverse. This study aimed to mine new molecular targets for the detection of Cronobacter strains at the species level. Using a bioinformatics approach and the online BLAST software, we obtained serial Cronobacter species-specific target genes. Primers for the selected genes were constructed, and the PCR products were evaluated using various Cronobacter species and non-Cronobacter strains. As a result, 2, 5, 7, and 5 specific target genes were found for C. sakazakii, C. malonaticus, C. dublinensis, and C. turicensis, respectively. Thereafter, 4- and 5-multiplex PCR (mPCR) was successfully developed to identify these four species based on the validated species-specific primers. Overall, our data indicated that PCR and mPCR assays have excellent specificity and sensitivity for the detection of Cronobacter species, suggesting the potential of their use in food inspection and clinical diagnosis.
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