A PCR-Colorimetric Microwell Plate Hybridization Assay for Detection of Mycobacterium tuberculosis and M. avium from Culture Samples and Ziehl-Neelsen-Positive Smears
| Autor: | Fabio Franzetti, Giulia Marchetti, Anna Degli Esposti, Andrea Gori, Chiara De Maddalena, Lidia Catozzi, Giulio Ferrario, Alessandra Bandera, Gianguglielmo Zehender, Maria Cristina Rossi |
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| Rok vydání: | 2000 |
| Předmět: |
DNA
Bacterial Microbiology (medical) Tuberculosis Lymph node biopsy Biology Polymerase Chain Reaction Specimen Handling Microbiology law.invention Mycobacterium tuberculosis Feces fluids and secretions law RNA Ribosomal 16S medicine Humans Polymerase chain reaction DNA Primers Mycobacterium Infections Suppuration medicine.diagnostic_test Sputum Mycobacteriology and Aerobic Actinomycetes Ribosomal RNA bacterial infections and mycoses 16S ribosomal RNA biology.organism_classification medicine.disease Virology Ziehl–Neelsen stain Colorimetry Lymph Nodes medicine.symptom Mycobacterium avium |
| Zdroj: | Journal of Clinical Microbiology. 38:1772-1776 |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.38.5.1772-1776.2000 |
| Popis: | Differentiation between Mycobacterium tuberculosis and M. avium is essential for the treatment of mycobacterial infections. We have developed an easy and rapid detection assay for the diagnosis of mycobacterial diseases. This is a PCR-hybridization assay based on selective amplification of a 16S rRNA gene sequence using pan- Mycobacterium primers followed by hybridization of the amplification products to biotinylated M. tuberculosis and M. avium -specific probes. A total of 55 mycobacterial isolates were tested. For all isolates, results concordant with those of conventional identification methods were obtained. Moreover, we developed a method for extraction of DNA from Ziehl-Neelsen-positive smears which allows the recovery of intact target DNA in our PCR-hybridization assay. Our method was able to confirm all culture results for 59 Ziehl-Neelsen-positive smears from clinical specimens (35 sputum, 11 lymph node biopsy, 6 stool, 4 pus, 2 urine, and 1 pericardial fluid specimens). These data suggest that our PCR-hybridization assay, which is simple to perform and less expensive than commercial probe methods, may be suitable for the identification of M. tuberculosis and M. avium . It could become a valuable alternative approach for the diagnosis of mycobacterial infections when applied directly to DNA extracted from Ziehl-Neelsen-positive smears as well. |
| Databáze: | OpenAIRE |
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