Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues
| Autor: | Maria Vitale, Yarelys Zamora Martínez, Carmen Fernandez Molina, Aleco D'Andrea, Rosa Alduina, V. Monteverde |
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| Přispěvatelé: | D'Andrea, A, Martinez, YZ, Alduina, R, Monteverde, V, Molina, CF, Vitale, M |
| Předmět: |
DNA
Bacterial Microbiology (medical) Serotype lcsh:Arctic medicine. Tropical medicine Tissue Fixation lcsh:RC955-962 lcsh:QR1-502 Kidney Settore BIO/19 - Microbiologia Generale Polymerase Chain Reaction lcsh:Microbiology law.invention 23S ribosomal RNA law Leptospira Formaldehyde RNA Ribosomal 16S medicine diagnostics Animals FFPE tissue Lung Polymerase chain reaction Paraffin Embedding biology microbiology Ribosomal RNA biology.organism_classification 16S ribosomal RNA medicine.disease Leptospirosis Molecular biology RNA Ribosomal 23S PCR Cattle Leptospira interrogans |
| Zdroj: | Scopus-Elsevier Memórias do Instituto Oswaldo Cruz, Volume: 107, Issue: 1, Pages: 85-88, Published: FEB 2012 Memórias do Instituto Oswaldo Cruz., Vol 107, Iss 1, Pp 85-88 (2012) |
| Popis: | In this study we compared two polymerase chain reaction (PCR) methods using either 16S ribosomal RNA (rRNA) or 23S rRNA gene primers for the detection of different Leptospira interrogans serovars. The performance of these two methods was assessed using DNA extracted from bovine tissues previously inoculated with several bacterial suspensions. PCR was performed on the same tissues before and after the formalin-fixed, paraffin-embedding procedure (FFPE tissues). The 23S rDNA PCR detected all fresh and FFPE positive tissues while the 16S rDNA-based protocol detected primarily the positive fresh tissues. Both methods are specific for pathogenic L. interrogans. The 23S-based PCR method successfully detected Leptospira in four dubious cases of human leptospirosis from archival tissue specimens and one leptospirosis-positive canine specimen. A sensitive method for leptospirosis identification in FFPE tissues would be a useful tool to screen histological specimen archives and gain a better assessment of human leptospirosis prevalence, especially in tropical countries, where large outbreaks can occur following the rainy season. |
| Databáze: | OpenAIRE |
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