Turbidity detection of shrimp taura syndrome virus by loop-mediated isothermal amplification reaction

Abstract

Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay is a novel method of gene amplification that amplifies nucleic acid, which can be applied for disease diagnosis in shrimp aquaculture. During the LAMP reaction, the white precipitate of magnesium pyrophosphate (Mg2P 2O7) is formed correlates with the amount of synthesized DNA. So, the turbidity can be measured. In this study, a portable turbidimeter has been developed for field to detection of Taura Syndrome Virus (TSV) that causes large economic losses to most major shrimp-producing countries including Thailand. The device could maintain an optimal temperature (63 °C) for 25 μl of LAMP sample solution contained in a 0.2 ml commercial PCR tube. We also applied the spectroscopic measurement technique to monitor a by-product of LAMP reaction, light emitting diode (LED) was used as a light source. Light dependent resistance (LDR) was used as detector. The results obtained from turbidity measurement revealed the same detection limit to those from agarose gel electrophoresis method. ©2010 IEEE.