Detection of Mycobacterium Bovis in Formalin-Fixed, Paraffin-Embedded Tissues of Cattle and Elk by PCR Amplification of an IS6110 Sequence Specific for Mycobacterium Tuberculosis Complex Organisms
| Autor: | Jack C. Rhyan, David L. Suarez, Janice M. Miller, Dennis A. Saari, Allen L. Jenny |
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| Rok vydání: | 1997 |
| Předmět: |
0301 basic medicine
Pathology medicine.medical_specialty Tuberculosis 040301 veterinary sciences Paratuberculosis Polymerase Chain Reaction Sensitivity and Specificity law.invention Microbiology 0403 veterinary science Mycobacterium tuberculosis 03 medical and health sciences law Formaldehyde medicine Animals Insertion sequence Polymerase chain reaction DNA Primers Mycobacterium bovis General Veterinary biology Deer 04 agricultural and veterinary sciences medicine.disease biology.organism_classification 030104 developmental biology Mycobacterium tuberculosis complex Paraffin DNA Transposable Elements Cattle Tuberculosis Bovine Mycobacterium |
| Zdroj: | Journal of Veterinary Diagnostic Investigation. 9:244-249 |
| ISSN: | 1943-4936 1040-6387 |
| DOI: | 10.1177/104063879700900304 |
| Popis: | A presumptive diagnosis of tuberculosis can be made if a tissue has characteristic histopathologic changes and acid-fast organisms. However, definitive diagnosis requires culture and species identification of the causative mycobacterium, a process that takes several weeks to complete. The purpose of work reported here was to determine if formalin-fixed, paraffin-embedded tissues could be tested by polymerase chain reaction (PCR) to provide a more rapid diagnosis of tuberculosis. Nondecalcified tissues from cases of tuberculosis in cattle and elk ( Cervus elaphus) were examined. The primers used for PCR amplified a 123-bp fragment of IS6110, an insertion sequence that is specific for organisms in the Mycobacterium tuberculosis complex ( M. tuberculosis, M. bovis, M. microti, M. africanum). The PCR test detected this sequence in tissues from 92 of 99 (93%) tuberculosis cases, including 3 of 4 elk. In 80 tissues, the positive results were obtained using material prepared by immersion of paraffin sections in water containing a detergent, followed by alternating boil/freeze cycles. The remaining positive results were obtained with DNA isolated from the crude tissue extracts by proteinase K digestion and phenol/chloroform purification. Accuracy of the IS6110 PCR test was demonstrated by negative test results on 31 tissues that had either nonmycobacterial granulomas or granulomatous lesions caused by other mycobacteria ( M. paratuberculosis or M. avium). The findings of this study show that a PCR test usually can provide a rapid diagnosis of tuberculosis when it is applied to paraffin sections that have characteristic lesions and acid-fast organisms. |
| Databáze: | OpenAIRE |
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