Comparison of four RNA isolating methods for identification of spring viraemia of carp virus (SVCV)

Authors

Abstract

Spring viraemia of carp virus (SVCV) , a negative sense single stranded RNA virus of the
family Rhabdoviridae, is the causative agent of a highly contagious SVC disease that
primarily affects the common carp ( Cyprinus carpio), an economically important fresh water
fish species with world-wide distribution.SVCV has also been reported to cause disease in
other fishes such as Poeciliidae, Esocida , Centrarchidae , Siluridae and salmonidae . There
are several diagnostic tests for the detection of SVC virus,however, the tests have not been
validated. The reverse transcriptase – polymerase chain reaction (RT-PCR) techniques have
been developed and validated representing a powerful tool for detection of RNA. One of the
most important aspects isolating RNA is to prevent degradation of the RNA during the
isolation procedure. In this study, we explored the efficiency of protocols for RNA isolation
from the SVCV strain 56/70.For RNA isolation, we compared four protocols, two guanidine
isotiocyanate phenol – chloroform based protocols ( RNX – Plus Iran , Iq2000 kit Taiwan )
and two column based protocols ( Cinnapure RNA Iran , high pure viral RNA kit , Roche
Germany ) that were commercially available. The results showed that the column based
protocols, Roche method and Cinapure performed better than other methods with the yields
of 31.76 ng/μl, 16/21 ng/μl, respectively. Each protocol yielded good quality of total RNA
bands (480 bp) being observed in agarose gel electrophoreses but was not observed in IQ2000
kit. Amount of total RNA isolated was lower for IQ2000 kit Protocol. Further, the RNA
being extracted from SVC by column based protocol method were resulted in successful
amplified using RT-PCR method

Keywords

Iranian Scientific Fisheries Journal
Volume 22, Issue 4 - Serial Number 85
November and December 2014
Pages 51-59

Files

Share

How to cite

Statistics