Detection of newly recognized rodent parvoviruses by PCR.
Several autonomous parvovirus isolates distinct from the prototypic rodent parvoviruses have recently been identified. These include variants of a mouse orphan parvovirus (MOPV) and a hamster isolate designated hamster orphan parvovirus (HOPV). In this study, a PCR primer set specific for these newly identified rodent parvoviruses was designed on the basis of DNA sequence comparisons of these isolates with other autonomous parvoviruses. The specificity of the primer set was determined by testing viral preparations of seven different parvoviruses and eight other viruses known to infect rodents. The PCR assay amplified the expected 260-bp product only in the presence of DNA from MOPV, HOPV, or LuIII a parvovirus of unknown species origin. The assay was able to detect as little as 10 pg of MOPV viral DNA or 1 pg of HOPV viral DNA, and it was able to detect MOPV in tissues from naturally infected mice and HOPV in tissues from experimentally infected hamsters. In contrast, the 260-bp product was not amplified from tissues of MOPV-negative mice or mock-infected hamsters. Our findings indicate that this PCR assay provides a rapid, specific, and sensitive method for the detection of MOPV in mice, HOPV in hamsters, and MOPV and HOPV in cell culture systems and that it may also be useful for the detection of LuIII contamination of cell culture systems.
Besselsen
D G
DG
Department of Veterinary Pathology, University of Missouri, Columbia 65211, USA.
Besch-Williford
C L
CL
Pintel
D J
DJ
Franklin
C L
CL
Hook
R R
RR
Riley
L K
LK
D000818:Animals; D002213:Capsid / Q000235:genetics; D006224:Cricetinae; D017931:DNA Primers; D004279:DNA, Viral / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D005260:Female; D007668:Kidney / Q000821:virology; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D017992:Parvovirus / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012376:Rodent Diseases / Q000821:virology*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004279:DNA, Viral