In situ hybridization法によるprogressive multifocal leukoencephalopathy剖検凍結脳におけるpapovaウイルス核酸の検索—ラジオアイソトープ標識probeとbiotin標識probeとの比較

THE DETECTION OF PAPOVAVIRUS NUCLEOTIDE SEQUENCES IN FROZEN BRAIN TISSUE FROM A PATIENT WITH PROGRESSIVE MULTIFOCAL LEUKOENCEPHALOPATHY BY IN SITU HYBRIDIZATION:COMPARISON OF IN SITU HYBRIDIZATION USING ISOTOPIC AND BIOTINYLATED PROBES

　抄録　³おH標識JCウイルスprobeとWardらによるbiotin標識JCウィルスprobeを用いてprogressive multifocal leukoencephalopathy （PML）凍結脳でin situ hybridizationを有ない，その有用性を比較した。³H標識法では長時間（3週間）の露出時間が必要であるがRigby法によりhigh specificactivityに標識されるのでsensitivityは高いと考えられた。しかし，biotin標識法ではhybridization後のhybridized部位の検出に用いるavidin-biotin peroxidase complexによる反応は数時間で行うことができ，さらにアイソトープの廃棄，検者の被曝の危険がないので，本法はウイルス性中枢神経感染症の診断法に応用できる可能性があると考えられた。

Ward et al developed biotinylated probes to hasten virus nucleic acid detection. We used these probes to trace SV 40 and JC virus nucleic acids in PML brain. This procedure visualizes the hy-bridization of viral sequences by affinity cytoche-mistry with avidin-biotin peroxidase complexes and diaminobenzidine. In the PML frozen white matter numerous oligodendroglial nuclei hybridi-zed with JC virus biotinylated and tritium label-led probes in areas adjacent to active demyelina-tion. Although tritium labeled probes was most sensitive and gave lowest background this bio-tinylated method showed be useful as a rapid diagnostic test for the specific detection of viral nucleic acid sequences in brain tissues from pati-ents with central nervous system (CNS) infectious diseases.