Abstrato

First Detection of Spring Viremia Of Carp Virus in Cyprinus Carpio, In Lake Shkodra/Scadar Albania

Berberi Enkeleda, Shumka Spase

Spring viremia of carp is a highly contagious viral disease caused by Rhabdovirus carpio which is a single strand RNA virus. A combination of one step RT-PCR (reverse transcription) and Semi-nested PCR was used to detect spring viremia of carp virus in infected cell cultures (EPC) and fish tissues. All the fish samples were collected from Lake Shkodra and from artificial ponds in different region of Albania. Total RNA was extracted from 500 μl of fish tissue extract or from 100 μl of supernatant from infected EPC cell cultures. One step RT-PCR is performed using a modification of the methods of Stone et al.(1) in a 15 μl reaction mix volume. We used SVCV F1 (primer forward) 5’- TCT-TGG-AGC-CAA-ATA-GCT-CAR*-R*TC-3’ and SVCV R2 ( primer reverse) 5’-AGA-TGG-TAT-GGACCC- CAA-TAC-ATH*-CAN*-CAY*-3 primers for the amplification of 714 bp fragment of SVCV cDNA (2). The reaction mix was subjected to thermal cycles (2). Amplified cDNA was analyzed in agarose gel electrophoresis. For the second round of amplification we used a semi-nested PCR with SVCV F1 5’-TCT-TGG-AGC-CAA-ATA-GCT-CAR*- R*TC-3’ and SVCV R4 5’-CTG-GGG-TTT-CCN*-CCT-CAA-AGY*-TGY*-3* primers (1). Amplified fragment cDNA (606 bp) is analyzed by agarose gel electrophoreses (2).The results of gel electrophoreses was positive for three fish samples (Cyprinus carpio) collected from Lake Shkodra. This is the first report of the detection of spring viremia of carp virus in Albania in a natural habitat like Lake Shkodra.