Development of recombinase polymerase amplification detection method for Listeria monocytogenes in food

Abstract

ObjectiveTo establish a recombinase polymerase amplification (RPA) method for the detection of Listeria monocytogenes (L. monocytogenes) in food.MethodsBased on the hlyA gene of L. monocytogenes, a set of RPA primers was selected for constructing RPA test, and its specificity and sensitivity were then tested.ResultsThe RPA assay could be finished in 30 min at 37 ℃. The primers used in RPA were specific. Experiments confirmed a detection limit of DNA template as low as 0.5 ng/μL. L. monocytogenes in the artificially-contaminated meat could be detected at the limit of 104 CFU/2.5 g.ConclusionThe RPA method for detecting L. monocytogenes has strong specificity and high sensitivity, which is easy to operate, and can be performed under normal temperature without expensive equipment. It is suitable for field detection and application in the basic laboratory.

Keywords

• listeria monocytogenes
• recombinase polymerase amplification
• hlya
• detection