Diagnostic polymerase chain reaction assays for identification of murine polyomaviruses in biological samples.
OBJECTIVE
Mouse polyoma virus and K virus are murine polyomaviruses frequently used in carcinogenicity and cellular biology studies in mice. These viruses can cause persistent infections, which increase the likelihood of transmission through transplantation of cells from infected mice. To identify polyomavirus-infected biological samples, several diagnostic polymerase chain reaction (PCR) assays were developed.


METHODS
Polyomavirus-family and virus-specific PCR assays were designed and optimized for specificity and sensitivity. The generic (polyomavirus-family) PCR assay and mouse polyoma virus-specific assays were compared with the mouse bioassay for diagnosis of infected cellular samples.


RESULTS
Specificity of the PCR assays was confirmed by testing a battery of other murine viruses. The mouse polyoma virus PCR test was the most sensitive assay, detecting as few as 2,000 copies of homologous virus. The K virus PCR assay was about eightfold less sensitive, and the generic PCR test was the least sensitive. Mouse polyoma virus and generic PCR assays amplified mouse polyoma virus in the inoculum and tissues from experimentally infected mice, and performed better than did the mouse bioassay.


CONCLUSIONS
Results of this study confirm that PCR is a specific and sensitive method for detection of murine polyomaviruses in biological samples.
Carty
A J
AJ
University Laboratory Animal Resources, University of Pennsylvania, Philadelphia 19104, USA.
Franklin
C L
CL
Riley
L K
LK
Besch-Williford
C
C
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007756:Laboratory Animal Science / Q000379:methods; D051379:Mice / Q000821:virology*; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D016133:Polymerase Chain Reaction / Q000379:methods*; D011120:Polyomavirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D027601:Polyomavirus Infections / Q000175:diagnosis / Q000662:veterinary*; D012680:Sensitivity and Specificity; D014412:Tumor Virus Infections / Q000175:diagnosis / Q000662:veterinary*
D000914:Antibodies, Viral; D004279:DNA, Viral