Rapid Detection of Norovirus GII by Fluorescent Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) and Nanomagnetic Bead Separation

Abstract

Noroviruses (NoVs) is the main cause of gastroenteritis in humans worldwide, mainly affecting school-age children and adults. NoVs are transmitted through feces and vomitus, including human contact, food, and water. Presently, NoVs are detected using molecular biological methods. Loop-mediated isothermal amplification (LAMP), specifically, requires little detection equipment, a short detection time, and low technical skills. Here, we established our own NoV reverse transcription (RT) polymerase chain reaction (PCR) quantitative detection system and a NoV GII RT-LAMP detection system. We collected 40 clinical samples, extracted RNAs, and used RT-PCR and RT-LAMP to detect NoV GII. The qualitative results of RT-LAMP were consistent with those of RT-PCR. However, a significant difference was observed between RT-LAMP and RT-PCR quantitative detection results. The NoV GII RT-LAMP detection system showed good sensitivity, up to 101, as well as good specificity. Furthermore, GI and GII did not interfere with each other. No false-positive responses were obtained for other gastrointestinal RNA viruses, such as Coxsackie virus A16 or enterovirus 71. Our results showed that the RT-LAMP detection system for NoV GII is suitable for the quantitative determination of NoV.