DEVELOPMENT OF A DUPLEX REALTIME PCR SYSTEM FOR THE SIMULTANEOUS DETECTION OF Hepatopancreatic parvovirus (HPV) AND Monodon baculovirus (MBV) IN SHRIMP

Abstract

Both MBV (Monodon baculovirus) and HPV Hepatopancreatic parvovirus) are shrimp enteric viruses that infect intestinal and hepatopancreatic epithelial cells to cause significant mortalities and depressed growth in infected larval, postlarval, and early juvenile stages of shrimp, and thus present a risk to commercial aquaculture. Therefore, in this study, a procedure for detecting of monodon baculovirus (MBV) and hepatopancreatic parvovirus (HPV) in shrimp by duplex realtime PCR assay using TaqMan probes  in a one-tube PCR reaction has been developed to quickly and accurately detect virus. The optimized duplex realtime PCR protocol include concentrations of primers HPVth/MBV 400 nM/400 nM, concentrations of probe HPV-P1/MBV-P1 là 300 nM/300 nM with the following conditions: initial denaturation at 95°C for 5 min, followed by 40 cycles of amplification at 95°C for 15s and annealing/extension at 60°C for 1 min. The results of evaluating of method show that the LAMP method have sensitivity (SE) is 100%, accuracy is (AC) 100%, specificity (SP) is100%, the false positive rate is 0 % and false negative rate is 0%.