EVALUATION OF NUCLEIC-ACID AMPLIFICATION METHODS FOR THE DETECTION OFHOG-CHOLERA VIRUS

A blind panel was tested in a diagnostic evaluation of a reverse trans cription (RT) polymerase chain reaction (PCR) method for detecting hog cholera virus (HCV) from pig tissues. The capability of the RT-PCR te st to discriminate between HCV and related pestiviruses, bovine viral diarrhea virus (BVDV), and those viruses causing similar diseases in s wine, including African swine fever virus (ASFV) and pseudorabies viru s (PRV), was also considered. Nucleic acid extraction involved either kit-based or conventional phenol:chloroform:isoamyl alcohol methods. A single-round PCR assay, using primers that hybridize to the conserved p120 nonstructural gene region, eras 82.5% sensitive (n = 17) and 100 % specific (n = 18) in the detection of the presence of HCV RNA. Howev er, the sensitivity was increased to 100% following a second PCR test. In all, 4 HCV, 7 BVDV, 2 ASFV, and 1 PRV isolates were studied. Novel nucleic acid sequences were generated for 9 HCV strains. Analysis of a portion of the p120 region using these methods was suitable for HCV isolate characterization.