Multiplex real-time PCR and high-resolution melting analysis for detection of white spot syndrome virus, yellow-head virus, and Penaeus monodon densovirus in penaeid shrimp

► The developed multiplex real-time PCR and HRM analysis for detection of PmDNV, WSSV, and YHV was highly specific and sensitive. ► Detection of the three viruses in forty-one shrimp samples demonstrated the applicability of this method to examine the viral infection in shrimp. ► The most attractive of this assay is its ability to simultaneously detect three of the major viruses, no requirement of probe and carcinogen dye, and capable to detect viral infection in cultured penaeid shrimp sample.