In situ hybridization of virulent canine distemper virus in brain tissue, using digoxigenin-labeled probes

Andreas Zurbriggen From the Institute of Animal Neurology, University of Berne, CH-3012 Berne, Switzerland.

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Cornelia Müller From the Institute of Animal Neurology, University of Berne, CH-3012 Berne, Switzerland.

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Marc Vandevelde From the Institute of Animal Neurology, University of Berne, CH-3012 Berne, Switzerland.

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Summary

Only a few hybridization experiments have been performed for detection of canine distemper virus (cdv) nucleic acid sequences in tissue cultures and in various tissues. Those published studies used probes derived from tissue culture-adapted cdv, and hybridization signals were not obtained in the CNS tissue, although infective cdv and viral antigen were detectable in this tissue. We developed probes complementary to virulent cdv and were able to detect viral RNA not only in primary brain cell cultures, but also in brain tissues, by use of in situ hybridization. Sensitivity of the test at least equaled that of immunohistochemistry. We applied digoxigenin-labeled, strand-specific RNA probes complementary to the nucleoprotein-coding viral nucleic acid sequence. Our results indicate that to detect cdv nucleic acid sequences in brain tissues, it is essential to use probes derived from the virulent virus.

Summary

Only a few hybridization experiments have been performed for detection of canine distemper virus (cdv) nucleic acid sequences in tissue cultures and in various tissues. Those published studies used probes derived from tissue culture-adapted cdv, and hybridization signals were not obtained in the CNS tissue, although infective cdv and viral antigen were detectable in this tissue. We developed probes complementary to virulent cdv and were able to detect viral RNA not only in primary brain cell cultures, but also in brain tissues, by use of in situ hybridization. Sensitivity of the test at least equaled that of immunohistochemistry. We applied digoxigenin-labeled, strand-specific RNA probes complementary to the nucleoprotein-coding viral nucleic acid sequence. Our results indicate that to detect cdv nucleic acid sequences in brain tissues, it is essential to use probes derived from the virulent virus.

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