The development of loop-mediated isothermal amplification combined with lateral flow dipstick for detection of Vibrio parahaemolyticus

• DOI: 10.1111/j.1472-765x.2011.03007.x
• Publication Date: 2011-01-18T20:31:21Z
• Authors: P. Prompamorn; P. Sithigorngul; S. Rukpratanporn; S. Longyant; P. Sridulyakul; P. Chaivisuthangkura
• Keywords: Thermolabile; Amplicon

The current study was aimed to develop a loop-mediated isothermal amplification (LAMP) combined with amplicon detection by chromatographic lateral flow dipstick (LFD) assay for rapid and specific of Vibrio parahaemolyticus.Biotinylated LAMP amplicons were produced set four designed primers that recognized specifically the V. parahaemolyticus thermolabile haemolysin (tlh) gene followed hybridization an FITC-labelled probe LFD detection. optimized time temperature conditions 90 min at 65 °C. LAMP-LFD method accurately identified 28 isolates but did not detect 24 non-parahaemolyticus 35 non-Vibrio bacterial isolates. sensitivity in pure cultures 120 CFU ml⁻¹. In case spiked shrimp samples without enrichment, limit 1·8 x 10³ g⁻¹ or equivalent 3 per reaction while conventional PCR 30 reaction.The established targeting tlh specific, sensitive identification parahaemolyticus.The developed provided valuable tool can be used effectively contaminated food sample.