Detection of Mycobacterium tuberculosis complex in paraffin-embedded tissue specimens
Journal Title: Postępy Nauk Medycznych - Year 2015, Vol 28, Issue 4
Abstract
Introduction. Molecular methods of diagnosis enabled the detection of M. tuberculosis complex directly in clinical materials, now make it possible also in the tissue embedded in paraffin blocks. Use of molecular methods to confirm the presence of Mycobacterium tuberculosis complex in paraffin-fixed material is particularly important for patients whose diagnosis based solely on the tissue and were not microbiologically tested.Aim. The aim of the research was to evaluate the usefulness of genetic methods to confirm the presence of Mycobacterium tuberculosis complex in clinical specimens on paraffin.Material and methods. 63 clinical specimens embedded in paraffin from patients with suspected tuberculosis. 1. Deparaffinization of specimens in xylene and dehydration in 96% ethanol by succeesive centrifugation. 2. Isolation DNA with the appropriate set to tissues embedded in paraffin blocks. 3. BD ProbeTec: The ampliflcation (in microwell) of Mycobacterium DNA, detection of the copied deoxyribonucleic acid in the process of the hybridization with species probe. Detected is insertion sequence IS6110 characteristic for the group M. tuberculosis complex. 4. GeneXpert Dx Test Xpert MTB/RIF is based on the reaction of nested real-time PCR. Test Xpert MTB/RIF is used to detect Mycobacterium tuberculosis and resistance to RMP.Results. The molecular analysis revealed the presence of M. tuberculosis complex DNA in 21 samples, whereas acid fast bacilli could be detected by Ziehl-Neelsen staining in only 13 samples. Z-N was negative in 41 of the patients, for 33 of them the result was compatible with the negative genetic test.Conclusions. Molecular methods are very important to complement and verify the microbiological and pathological results.
Authors and Affiliations
Sylwia Brzezińska, Małgorzata Szołkowska, Renata Langfort, Zofia Zwolska, Ewa Augustynowicz-Kopeć
Keywords
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