title: Development and evaluation of a RT-LAMP assay for rapid detection of hepatitis E virus from shellfish.
abstract: 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 °C) for 60 min, and can be visually evaluated by staining at a time in about 1h. 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.
authors.affiliation: College of Animal Husbandry & Veterinary Medicine, Liaoning Medical University, No. 5-48 Renmin Street, Jinzhou 121001, China; College of Food Science, Shenyang Agricultural University, No. 120 Dongling Road, Shenyang 110866, China. Electronic address: gordon402@163.com.
authors.forename: Shenyang
authors.initials: S
authors.lastname: Gao
authors.affiliation: Animal Quarantine Lab, Inspection & Quarantine Technology Center of Hainan Entry-Exit Inspection & Quarantine Bureau, Haikou 570000, China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Li
authors.affiliation: College of Animal Husbandry & Veterinary Medicine, Liaoning Medical University, No. 5-48 Renmin Street, Jinzhou 121001, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.affiliation: College of Animal Husbandry & Veterinary Medicine, Liaoning Medical University, No. 5-48 Renmin Street, Jinzhou 121001, China.
authors.forename: Enhui
authors.initials: E
authors.lastname: Zha
authors.affiliation: College of Animal Husbandry & Veterinary Medicine, Liaoning Medical University, No. 5-48 Renmin Street, Jinzhou 121001, China.
authors.forename: Shen
authors.initials: S
authors.lastname: Wang
authors.affiliation: College of Animal Husbandry & Veterinary Medicine, Liaoning Medical University, No. 5-48 Renmin Street, Jinzhou 121001, China.
authors.forename: Yonggang
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Animal Husbandry & Veterinary Medicine, Liaoning Medical University, No. 5-48 Renmin Street, Jinzhou 121001, China.
authors.forename: Tiezhong
authors.initials: T
authors.lastname: Zhou
authors.affiliation: College of Food Science, Shenyang Agricultural University, No. 120 Dongling Road, Shenyang 110866, China.
authors.forename: Xiqing
authors.initials: X
authors.lastname: Yue
mesh_terms: D000818:Animals; D002681:China; D005516:Food Microbiology / Q000379:methods*; D016752:Hepatitis E virus / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012758:Shellfish / Q000821:virology*; D014780:Viruses / Q000235:genetics
keywords: Hepatitis E virus; RT-LAMP; RT-qPCR; Shellfish