Development and evaluation of a RT-LAMP assay for rapid detection of hepatitis E virus from shellfish

摘要

Hepatitis E virus (HEV) has becoming a well known zoonotic enteric pathogen and circulated widely inter human-animal-water-food. Generally, detection of the virus has relied on conventional reverse transcription-PCR (RT-PCR) and TaqMan/SYBR quantitative real-time RT-PCR (RT-qPCR), but these tools are usually disadvantages in time-consuming and expensive instruments required. In the present study, we report here on the development of a one-step single-tube reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for rapid detection of HEV contamination in shellfish. The amplification is completed under the isothermal condition (63 degrees C) for 60 min, and can be visually evaluated by staining at a time in about 1 h. In addition, a total of 315 shellfish (80 Anadara granosa, 115 Scapharca subcrenata and 120 Ruditapes philippinarum) collected monthly from the Jinzhou coastal estuary of China Bohai gulf were investigated for HEV contamination by the RT-LAMP compared with a standard RT-qPCR. It was found that genotype 4 HEV was detected in all three species of shellfish sampled using the RT-LAMP assay and was in accordance with RT-qPCR detection of HEV in shellfish. Summarily, our results indicate that the RT-LAMP is a rapid, specific, sensitive and reliable method. This method offers a new tool for the routine monitoring of HEV contamination in shellfish or its harvesting waters in field. (C) 2015 Elsevier B.V. All rights reserved.