Development of one-step reverse transcription loop-mediated isothermal amplification for norovirus detection in oysters

Abstract

The aim of this study was to develop a simple and rapid technique for detecting human norovirus (HuNoV). The loop-mediated isothermal amplification (LAMP) technique was evaluated and found to be sensitive, highly specific, and useful for routine oyster testing. Reverse transcription-LAMP (RT-LAMP) primers were designed to detect HuNoVs genogroups I (GI) and GII. One-step RT-LAMP was found to be more sensitive than real-time RT-PCR, semi-nested RT-PCR and real-time RT-LAMP methods and could detect 101 HuNoV genome copies per microliter. The specificity of one-step RT-LAMP for HuNoVs GI and GII was confirmed by restriction fragment length polymorphism analysis. When a field test of oyster samples was performed, one-step RT-LAMP proved to be a rapid and accurate technique for detecting HuNoV. Thus, this could be reliable rapid technique for detecting HuNoV in fishery production fields, so to improve food safety and enable the oyster industry to become more competitive in the world market. © 2016 Elsevier Ltd