Comparison of Efficiency of Different Nucleic Acid Extraction Methods for Bovine Enteroviruses in Feces

Abstract

Bovine enteroviruses are thought to be mild pathogenic or non pathogenic viral gastroenteretis agents. However there are some cases where they are identified from cattle having different symptoms. They can also be used as markers of environmental contamination. Enterovirus family has fast replication capacity and can be used as potential vectors. For this purpose isolation and molecular studies concerning Enteroviruses are currently increasing. In this study 10 cattle feces samples were used for evaluation of 4 different extraction methods for investigation of bovine enteroviruses. These methods comprise of TRIzol LS (Life Technologies), QIAamp Viral RNA mini kit (Qiagen), NucleoSpin RNA (Macherey-Nagel) and phenol-chloroform extraction. All procedures performed in BSL-2 cabinet and RNA yield of extracts were analysed in Bio-spec (Shimadzu) spectrophotometer. This RNA templates were then tested by Verso 1 step RT PCR (Thermo Scientific) kit with spesific for Enterovirus 5’UTR primers. And observed for band formation. While QIAmp Viral RNA kit had the shortest protocol, it did not have as much sensitivity as TRIzol LS and TRIzol was determined as the most sensitive method to make extraction of bovine enteroviruses from feces.

Keywords

• Bovine Enteroviruses, RT-PCR, Extraction