Clinical Microbiology: Open Access

Rapid detection of Vibrio parahaemolyticus in oyster via loop-mediated isothermal amplification assay

3^rd International Conference on Clinical Microbiology & Microbial Genomics

September 24-26, 2014 Valencia Convention Centre, Spain

Chamjuree Malaivichit, Tanawut Tantimongcolwat, Srisurang Tantimavanich, Rungrot Cherdtrakulkiat and Virapong Prachayasittikul

Posters: Clin Microbial

Abstract:

Vibrio parahaemolyticus is found in the highest incident among pathogens associated with seafood-borne disease. It causes acute gastroenteritis in humans when ingestion of live pathogen. Therefore, early detection of V. parahaemolyticus is very important in order to prevent sickness caused by consumption of contaminated seafood. The reference method for detection of V. parahaemolyticus is its culture and biochemical test. Unfortunately, such method is labor-intensive and timeconsuming. Thus, it is developed a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of V. parahaemolyticus in oyster sample. The optimal conditions for LAMP reaction were found to be 2 mM MgSO4, 1.2 mMdNTPs and 0.2 M betaine at 65?C for 30 min. It was able to detect genomic DNA of V. parahaemolyticus as low as to 1 fg/μl. The assay showed 100% detection accuracy to V. parahaemolyticus against 6 other species of Vibrio and 17 non-Vibrio spp. For spiked oyster sample without enrichment process, the detection limit of V. parahaemolyticus was 100cfu/g. Therefore, the LAMP assay developed in this study provided high sensitivity and specificity of detection toward V. parahaemolyticus with a great potential for future field detection of V. parahaemolyticus in raw seafood samples.

Biography :

Chamjuree Malaivichit is an MSc student at the Faculty of Medical Technology, Mahidol University, Thailand. Now, she is challenged with the development of technology for rapid screening of V. parahaemolyticusin order to ensure food safety.