Evaluation of nucleic acid amplification methods for the detection of hog cholera virus.
A blind panel was tested in a diagnostic evaluation of a reverse transcription (RT) polymerase chain reaction (PCR) method for detecting hog cholera virus (HCV) from pig tissues. The capability of the RT-PCR test to discriminate between HCV and related pestiviruses, bovine viral diarrhea virus (BVDV), and those viruses causing similar diseases in swine, including African swine fever virus (ASFV) and pseudorabies virus (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, was 82.5% sensitive (n = 17) and 100% specific (n = 18) in the detection of the presence of HCV RNA. However, 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.
Harding
M J
MJ
Animal Diseases Research Institute, Ontario, Canada.
Prud'homme
I
I
Gradil
C M
CM
Heckert
R A
RA
Riva
J
J
McLaurin
R
R
Dulac
G C
GC
Vydelingum
S
S
D000358:African Swine Fever Virus / Q000302:isolation & purification; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D006691:Classical Swine Fever / Q000175:diagnosis*; D006692:Classical Swine Fever Virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017124:Conserved Sequence; D017931:DNA Primers; D011558:Herpesvirus 1, Suid / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010576:Pestivirus / Q000302:isolation & purification*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D013552:Swine; D017361:Viral Nonstructural Proteins / Q000235:genetics
D017931:DNA Primers; D011933:Reagent Kits, Diagnostic; D017361:Viral Nonstructural Proteins