|Jeon, Su Been - Chung-ang University|
|Seo, Dong Joo - Chung-ang University|
|Oh, Hyejin - Chung-ang University|
|Choi, Changsun - Chung-ang University|
Submitted to: Food Control
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/2/2016
Publication Date: 3/5/2017
Citation: Jeon, S., Seo, D., Oh, H., Kingsley, D.H., Choi, C. 2017. Development of one-step Loop-Mediated Isothermal Amplification (LAMP) for the detection of norovirus in oysters. Food Control. 73:1002-1009.
Interpretive Summary: Human norovirus is the number one cause of foodborne illness and is a worldwide problem for the shellfish industry. Current detection technologies can quantitate norovirus within shellfish tissues but these techniques are too cumbersome for routine market shellfish samples in a field setting. In this publication, a simple isothermal method is successfully applied to oyster testing. This detection technique requires only a heat block or temperature-controlled water bath and positives samples can be assessed visually by test tube color changes. The new method was found to be more sensitive than conventional methods and it was also was readily able to pick out the pathogen among other organisms present. In a field test with oysters, the new method proved to be rapid and accurate. Results indicate that this technique should facilitate routine field testing for norovirus within shellfish.
Technical Abstract: The aim of this study was to develop a simple and rapid technique for detecting human norovirus (NoV). 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 norovirus genogroups GI and GII. One-step RT-LAMP was found to be more sensitive than real-time RT-PCR and semi-nested RT-PCR methods and could detect 101 NoV genome copies per microliter. The specificity of one-step RT-LAMP for NoV 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 NoV. This technique could thus be used to improve food safety and enable the oyster industry to become more competitive in the world market.