A Loop-Mediated Isothermal Amplification Method for the Detection of Human Enterovirus in Food Packaging Film Samples

摘要

In order to reduce food-borne outbreaks, detection must be taken at all points in the farm-to-tablechain including packing. Human enteroviruses (HEVs) widely exist in most environments that may cause aseries of diseases in humans. Rapid methods for the detection of HEVs in food packing process are importantto the food industry and for public health. The LAMP assay of HEVs reported in this study was highly specific,sensitive, and easily diagnostic. A set of four primers was specially designed to strictly detect HEVspreferably HEV-A and HEV-B. The amplification was carried out under isothermal conditions at 61 °C, andthe result could be obtained in less than 90 min and be monitored by naked-eye visualization. We observedspecifically only for HEVs that HEV-A and HEV-B like and not for other viruses. The sensitivity of the assaywas 101 copies/μl of a cloned enterovirus 71 fragment, which was equivalent to the sensitivity of the PCR.We tested the LAMP assay in different samples including packaging film samples using for ready-to-eat food,source water and stool specimens by using this LAMP system. A negative result showed in packaging filmwhile positive reaction could be observed in environmental samples and clinical specimens. These results indicatethat the developed LAMP assay here has the potential to become a practical method for the rapid diagnosingHEVs preferably HEV-A and HEV-B in various samples.