A METHOD FOR DIRECT DNA AMPLIFICATION OF UNCHARACTERIZED DNA VIRUSES AND FOR DEVELOPMENT OF A VIRAL POLYMERASE CHAIN-REACTION ASSAY - APPLICATION TO THE RED-SEA BREAM IRIDOVIRUS

A method is described for isolating a DNA segment of a virus for which no protein or DNA sequence information is available, This segment can then be used to develop a PCR-based assay for the virus. The method i s based on the widespread presence and strong conservation of the ribo nucleotide reductase gene among DNA viruses. The validity of the proce dure is demonstrated by development of an assay for the fish iridoviru s, We report the direct isolation from infected fish of a 738-bp segme nt of the iridovirus ribonucleotide reductase small subunit gene witho ut prior virus purification, Using the sequence information obtained, a PCR-based diagnostic system was developed for detecting iridovirus i nfection. (C) 1996 Academic Press, Inc.