A novel combination of TaqMan RT-PCR and a suspension microarray assay for the detection and species identification of pestiviruses.
The genus pestivirus contains four recognized species: classical swine fever virus, border disease virus, bovine viral diarrhoea virus types 1 and 2. All are economically important and globally distributed but classical swine fever is the most serious, concerning losses and control measures. It affects both domestic pigs and wild boars. Outbreaks of this disease in domestic pigs call for the most serious measures of disease control, including a stamping out policy in Europe. Since all the members of the pestivirus genus can infect swine, differential diagnosis using traditional methods poses some problems. Antibody tests may lack specificity due to cross-reactions, antigen capture ELISAs may have low sensitivity, and virus isolation may take several days or even longer time to complete. PCR-based tests overcome these problems for the most part, but in general lack the multiplexing capability to detect and differentiate all the pestiviruses simultaneously. The assay platform described here addresses all of these issues by combining the advantages of real-time PCR with the multiplexing capability of microarray technology. The platform includes a TaqMan real-time PCR designed for the universal detection of pestiviruses and a microarray assay that can use the amplicons produced in the real-time PCR to identify the specific pestivirus.
Joint R&D Division, Department of Virology, Immunobiology and Parasitology, National Veterinary Institute (SVA) and Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences (SLU), SE-751 89 Uppsala, Sweden. neil.leblanc@sva.se
Neil
N
LeBlanc
Mikael
M
Leijon
Magnus
M
Jobs
Jonas
J
Blomberg
Sándor
S
Belák
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D046228:Microarray Analysis / Q000379:methods*; D010576:Pestivirus / Q000145:classification* / Q000235:genetics*; D018182:Pestivirus Infections / Q000662:veterinary* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000821:virology*
D017931:DNA Primers