Duplex PCR protocol for the detection of Vibrio parahaemolyticus (VpAHPND) and White Spot Syndrome Virus (WSSV) causing diseases in marine shrimp

Abstract

The intensification of farming areas and climate change have made the shrimp farming industry suffer from various obstacles, especially the loss of productivity due to pathogens. White Spot Syndrome Virus (WSSV) and Vibrio parahaemolyticus (Vp_AHPND) were reported to cause huge economic losses to marine shrimp farmers . When causing diseases or outbreaks, they are extremely hard to be controlled. Therefore, detecting and screening out the pathogens are crucial. This study was conducted to optimize a duplex PCR (dPCR) protocol, which is able to detect WSSV and Vp_AHPND simultaneously. Benefits of the protocol are noticeable, such as resulting fast, and reducing the concentration of primer, dNTPs, and MgCl₂ while providing substantial sensitivity and specificity. The results showed that (i) The PCR protocol was successfully optimized to detect WSSV and Vp_AHPND simultaneously and its specificity was determined. (ii) For the DNA template of WSSV and V. parahaemolyticus, the protocol required 100 pg/µL and 10 ng/µl, respectively. (iii) The duplex PCR was successfully applied to detect WSSV and V. parahaemolyticus in marine shrimp samples.