Screening of specific nucleic acid targets for Cronobacter sakazakii and visual detection by loop-mediated isothermal amplification and lateral flow dipstick method in powdered infant formula

Due to the lack of specific genes for rapid detection methods Cronobacter sakazakii in food samples, whole genome sequence analysis was performed this investigation using basic local alignment search tool. Forty-two DNA fragments unique C. were mined, then primers designed and screened by PCR loop-mediated isothermal amplification (LAMP). Two primer sets, CS1 CS31, found as stable primers, with their corresponding nucleic acid targets CSK29544_00235 gene CSK29544_03484 gene, respectively. Furthermore, compared 3 reported previously, these 2 verified more among species, similarity MLST databases (http://pubmlst.org/cronobacter). The specificity LAMP reaction approached 100% 48 bacterial strains, which included 22 strains. Subsequently, combined visual lateral flow dipstick (LFD) based on above targets, demonstrated a rapid, efficient method high specificity. Finally, sensitivity assay system pure cultures artificially contaminated milk measured 4.5 × 100 cfu/mL 5.7 101 cfu/g, Total time within h. Thus, establishment LAMP-LFD shows great significance potential powdered infant formula.