Aquaculture & Fisheries

Biography

Biography: Wansika Kiatpathomchai

Abstract

An accelerated, loop-mediated isothermal amplification (LAMP) platform has been developed for sensitive and specific detection of few copies of target nucleic acids of pathogens. It operates under isothermal conditions by self-recurring strand-displacement DNA synthesis using specially designed primer sets. Although LAMP assays have been widely used in animal and human disease diagnosis, detection of positive LAMP reaction was conventionally visualized by agarose gel electrophoresis of amplicons with time consuming detection step. This can be avoided by replacement with cheaper, equally sensitive, more rapid and simpler methods including the use of a real time turbidimeter, use of lateral flow dipsticks or oligonucleotide-labled nanogold probes  and use of metal indicators. Since these LAMP platforms do not require sophisticated equipment, they are more applicable for small or field laboratories. The time required for these integrated LAMP platforms is approximately 70 min (10 min for DNA/RNA preparation, 30-45 min for the LAMP reaction and 10-15 min for detection of the positive LAMP result). These platforms have been successfully applied for detection of shrimp pathogens (i.e., white spot syndrome virus, yellow head virus, Taura syndrome virus, Laem Singh slow growth virus, microsporidia and acute hepatopancreatic necrosis disease) and fish (tilapia) pathogens (Streptococcus iniae, S. agalactiae, Flavobacterium columnare, Francisella noatunensis subsp. orientalis, Aeromonas hydrophila, Shewanella putrefaciens and Infectious Spleen and Kidney Necrosis Virus). These integrated LAMP platforms are sensitive, simple, rapid and cost-effective and can applied in routine detection and are also more suitable for field conditions making them more suitable for use in quarantine/surveillance programs