Development and Application of a Triplex PCR Assay for Detection of Koi Herpes Virus (KHV) Gene Infected Common Carp (Cyprinus carpio L.)

Abstract Koi herpesvirus (KHV) has been responsible for worldwide KHV disease (KHVD) outbreaks, resulting in high mortality rates, and reported Cyprinus carpio cultivation. A Triplex Polymerase Chain Reaction (T-PCR) was developed the detection of gene to improve diagnosis infection. Three sets primers were designed targeting specific sequences which frequently used PCR assay detecting (modifications primer by Yuasa,290 bp; Gilad 484 bp, TK, 409 bp) once running. The validity test conducted verified using DNA sample Aeromonas hydrophila . Sensitivity singleplex dilution. result showed that T-PCR method these pairs is at 55 annealing temperature with consistency repeated triplicates, same results. found no band detected A. This PCR-based may have useful clinical tool disease.