Detection of rodent parvoviruses by use of fluorogenic nuclease polymerasechain reaction assays

Polymerase chain reaction (PCR) assays have proven useful for detection of rodent parvoviruses in animals and contaminated biological materials. Fluor ogenic nuclease PCR assays combine PCR with an internal fluorogenic hybridi zation probe, eliminating post-PCR processing and potentially enhancing spe cificity. Consequently, three fluorogenic nuclease PCR assays were develope d, one that detects all rodent parvoviruses, one that specifically detects minute virus of mice (MVM), and one that specifically detects mouse parvovi rus 1 (MPV) and hamster parvovirus (HaPV). When rodent parvoviruses and oth er rodent DNA viruses were evaluated, the rodent parvovirus assay detected only rodent parvovirus isolates, whereas the MVM and MPV/HaPV assays detect ed only the MVM or MPV/HaPV isolates, respectively. Each assay detected the equivalent of 10 or fewer copies of target template, and all fluorogenic n uclease PCR assays exceeded the sensitivities associated with previously re ported PCR assays and mouse antibody production testing. In addition, each fluorogenic nuclease PCR assay detected the targeted parvovirus DNA in tiss ues obtained from mice experimentally infected with MVM or MPV. Results of these studies indicate that fluorogenic nuclease PCR assays provide a poten tially high-throughput, PCR-based method to detect rodent parvoviruses in i nfected mice and contaminated biological materials.