DETECTION OF CANINE-DISTEMPER VIRUS IN 100-PERCENT OF PAGETS-DISEASE SAMPLES BY IN-SITU REVERSE TRANSCRIPTASE-POLYMERASE CHAIN-REACTION

Previous evidence implicating paramyxoviruses in the aetiopathology of Paget's disease of bone has been controversial. While several groups have demonstrated the presence of paramyxoviruses using electron micro scopy, immunohistochemistry, and molecular biological techniques, othe rs have found no evidence of viruses using reverse transcriptase-polym erase chain reaction (RT-FCR), We have previously provided evidence th at canine distemper virus (CDV) is present in approximately 65% of sam ples of pagetic bone, using in situ hybridization and RT-PCR; however, these results have been criticized, To further investigate the possib le Role of CDV, we have now developed the technique of in situ-RT-PCR (IS-RT-PCR) to examine for the presence of CDV-nucleocapsid (CDV-N) ri bonucleic acid (RNA) in pagetic bone, Control samples consisted of uni nvolved sites from patients with the disease, normal bone, and several active remodeling states. IS-RT-PCR was optimized to detect CDV-N usi ng distemper-infected vero cells, The specificity of the technique was confirmed using vero cells infected with CDV, which showed amplified signal following IS-RT-PCR, and cells infected with measles virus (MV) , in which no positive signal for CDV was detected by IS-RT-PCR, Follo wing conventional in situ hybridization, CDV-N was detectable in 10 of 15 pagetic bone samples. However, after five, and particularly 10, cy cles of IS-RT-PCR, CDV-N was found in all 15 samples. There was no evi dence of CDV in four samples from uninvolved sites from pagetic patien ts, or in any of the other control samples. In this study, using the n ovel technique of IS-KT-PCR, CDV was found to be present in 100% of pa getic samples examined. There was no evidence of the virus in any of t he control samples, including samples of bone from uninvolved sites fr om patients with Paget's disease, These results provide additional pro of that CDV is present within pagetic bone and further support the hyp othesis that paramyxoviruses are involved in the etiopathology of Page t's disease, (Bone 23:171-175; 1998) (C) 1998 by Elsevier Science Inc. All rights reserved.