Immuno-enrichment and rapid detection of Enterobacter sakazakii

Objective To study a rapid detection method for Enterobacter sakazakii by combining immuno-enrichment with PCR.Methods PCR tubes were coated with monoclonal antibody against Enterobacter sakazakii to capture the suspected bacteria in the samples.PCR was carried out to detect the captured bacteria.The feasibility of the method was assessed with specificity,sensitivity,simulation enrichment experiments and comparison to the results of enzyme linked immunosorbent assay(ELISA).Results The sensitivity of this method was up to 10²-10³ cfu/mL.Compared to the results of direct PCR,the sensitivity of this method was increased by 10³-10⁴ times.No positive results was observed for 10 food-borne pathogens other than Enterobacter sakazakii in the tests.The sensitivity of this method in simulation enrichment experiment was up to 10³ cfu/mL.The method just needs 6 hours enrichment time,while ELISA method needed 16 hours enrichment time.Conclusion The method is sensitivie,specific,and rapid for the detection of Enterobacter sakazakii.