DEVELOPMENT OF PRIMERS FOR DETECTION OF HEPATOPANCREATIC PARVOVIRUS

摘要

Polymerase chain reaction (PCR) primers for detection of Hepatopancreatic parvo virus (HPV) in shrimp Shrimp farming is one of the sustainable activities having a stake in the coastal zone. Hepatopancreatic parvo virus (HPV) infects the cultured shrimp and causes stunted growth, which ultimately results in decreased production in shrimp farming. As there is no treatment for this disease, it is advised to screen the shrimp larvae before stocking in the production ponds. In order to undertake screening and earlier diagnosis of the disease caused by HPV, the Vaccine Research Centre-Viral Vaccines, Centre for Animal Health Studies, Tamil Nadu Veterinary and Animal Sciences University, has developed Polymerase chain reaction (PCR) primers for detection of Hepatopancreatic parvo virus (HPV) in shrimp. The PCR primers for nested PCR assay consist of two sets of primers. A pair of outer primers (SDDL-HPV F1 and SDDL-HPV R1) that amplify the portion of capsid protein gene of HPV yielding a product of 429 bp size upon amplification by PCR. A pair of inner (nested) primers (SDDL-HPV F2 and SDDL-HPV R2) that amplify a portion of capsid protein gene of HPV internal to the region amplified by the outer primers yielding a product of 188 bp size. The PCR primers for single step PCR consist of one set of primers (SDDL-HPV DGF1 and SDDL-HPV DGR1) that amplify a portion of capsid protein gene of two different HPV strains infecting the cultured shrimp belonging to species Penaeus monodon and Penaeus chinensis yielding a product of 397 bp size. The above primers were tested in shrimp samples collected from shrimp hatcheries and farms and were found to have a good sensitivity and specificity in detecting Hepatopancreatic parvo virus (HPV). These primers can be used for screening / diagnosis of HPV in larvae, postlarvae, juveniles, adults and broodstocks of shrimp which helps in prevention of the disease.