A New Validated Real-Time PCR-Based Method for the Specific and Fast Detection of Cronobacter spp. in Infant Formula

作者：Soler Marcal; Ruiz Rueda Olaya; Lopez Siles Mireia; Calvo Laia^*; Kaclikova Eva; Garcia Gil Jesus L

来源：Food Analytical Methods, 2012, 5(2): 179-187.

DOI：10.1007/s12161-011-9252-y

• Cronobacter spp.
• Enterobacter sakazakii
• Powdered infant formula (PIF)
• Real-time PCR
• Validation

In this study, a real-time polymerase chain reaction (PCR)-based method was designed for the fast detection of Cronobacter spp. (a newly proposed genus formerly known as Enterobacter sakazakii) in infant formula. The real-time PCR was positively tested with 70 Cronobacter strains, including members of all the species of this genus, and 88 non-Cronobacter strains. This new PCR-based system was validated against the reference standard ISO/TS 22964: 2006 (ISO International Organization for Standardization 2006) using 70 food matrices including powdered infant formula, follow-up formula, and hydrolyzed cereals for infants. The detection limit of the technique was found to be of 1 cfu in 10 g of food, fulfilling the requirements of the European Commission. The time of analysis, which comprises around 3-6 days using the reference method, is considerably reduced to less than 24 h using the real-time PCR-based system hereby described, allowing food industry a faster release of the stocks for commercialization. Moreover, this method includes an internal amplification control, co-amplified during each PCR run to verify the results.

• 出版日期2012-4