Fast short-fragment PCR for rapid and sensitive detection of shrimp viruses

This article describes a fast short-fragment PCR method for the detection of white spot syndrome virus (WSSV), infectious hypodermal and hematopoietic necrosis virus (IHHNV), and monodon baculovirus (MBV). Fast two-temperature (95 °C denaturation and 60 °C annealing/extension) PCRs were performed in 5–10 μl volume samples in miniaturized microplates using a fast Peltier thermal cycler. 40 cycles were completed in 25–30 min. Rapid high-resolution agarose gel electrophoresis of 70–150 bp PCR fragments was performed in 10 min. High sensitivity of PCR product detection (50–100 pg) was obtained using ultra sensitive dyes such as GelStar® and a gel documentation system equipped with a blue-light transilluminator. This novel method is faster and more sensitive than its TaqMan® real-time PCR counterparts.