Use of discriminatory probes for strain typing of formalin-fixed, rabies virus-infected tissues by in situ hybridization
| Autor: | Susan A. Nadin-Davis, Mary Sheen, Alexander I. Wandeler |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Microbiology (medical)
Tissue Fixation medicine.disease_cause Virus Mice Species Specificity Virology medicine Animals Typing Mononegavirales Lyssavirus In Situ Hybridization Retrospective Studies biology Reverse Transcriptase Polymerase Chain Reaction Rabies virus Genetic Variation RNA Probes Amplicon Rhabdoviridae biology.organism_classification medicine.disease Rabies |
| Zdroj: | Journal of clinical microbiology. 41(9) |
| ISSN: | 0095-1137 |
| Popis: | An in situ hybridization (ISH) method has been developed to overcome difficulties encountered in the viral typing of formalin-fixed rabies virus-infected brain tissue. Rabies viruses representative of all strains normally encountered in diagnostic submissions throughout Canada, including 3 strains of terrestrial hosts (arctic fox, western skunk, mid-Atlantic raccoon), 10 strains circulating in several bat reservoirs (BBCAN1 to BBCAN7, LACAN, SHCAN, and MYCAN), and the Evelyn-Rokitniki-Abelseth (ERA) strain, used as an oral vaccine for fox rabies control in Ontario, were targeted. Partial phosphoprotein gene fragments generated from reverse transcription (RT)-PCR products of specimens of each viral type were molecularly cloned and used to produce negative-sense digoxigenin-labeled RNA transcripts. Conditions permitting the use of these transcripts as strain-specific probes were optimized by blotting analyses with RT-PCR amplicons generated with representative rabies viruses and by ISH applied to mouse brains inoculated with these strains. The successful application of this methodology to two rabies virus-positive specimens that were also identified by traditional methods and the retrospective typing of two archival rabies virus-positive equine specimens is described. This technique provides a typing regimen for rabies virus isolates submitted in a form that is normally recalcitrant to alternate typing strategies. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|