DETECTION AND DIFFERENTIATION OF NON-TUBERCULOUS MYCOBACTERIA AND M. TUBERCULOSIS COMPLEX BY REAL TIME PCR
https://doi.org/10.21292/2075-1230-2016-94-9-80-87
Abstract
Goal of the study: to define the design of primers and probes specific to DNA of non-tuberculous mycobacteria and evaluate their diagnostic value in case of simultaneous detection of non-tuberculous mycobacteria and M. tuberculosis complex by real time PCR.
Materials and methods. Primer 3, Primer BLAST, Ugene Uni Pro were used to design primers and probes. Preliminary assessment of specificity and sensitivity of detection of non-tuberculous mycobacteria DNA was performed on cultures belonging to 18 types of non-tuberculous mycobacteria, 16 strains of M. tuberculosis complex and 14 types of microorganisms being none Mycobacterum. Analytic sensitivity was tested on 284 cultures of non-tuberculous mycobacteria and diagnostic sensitivity was tested on 124 sputum samples. The kit ofM-Sorb-Tub-Avtomat (ZAO Sintol) was used for DNA isolation. Cultures were subcultured on the liquid medium of Middlebrook 7H9 in Bactec MGIT 960. Cultures were identified with the use of standard microbiological techniques. Analysis of DNA isolated from cultures was performed by the reagent kit of GenoTypeCM/AS (Hain Lifescience, Germany).
Results. 100% specificity and sensitivity of PCR was demonstrated in mycobacterial cultures and 100% specificity and 69-70% sensitivity was demonstrated in diagnostic material analysis.
About the Authors
V. V. Ustinova
Central Tuberculosis Research Institute
Russian Federation
Russian Federation
Junior Researcher of Microbiological Department,
2, Yauzskaya alleya, Moscow, 107564
T. G. Smirnova
Central Tuberculosis Research Institute
Russian Federation
Russian Federation
senior Researcher of Microbiological Department,
2, Yauzskaya alleya, Moscow, 107564
D. A. Varlamov
All-Russia Research Institute of Agricultural Biotechnology
Russian Federation
Russian Federation
Researcher,
42, Timiryazevskaya st., Moscow, 127550
I. Yu. Andrievskaya
Central Tuberculosis Research Institute
Russian Federation
Russian Federation
Junior Researcher of Microbiological Department,
2, Yauzskaya alleya, Moscow, 107564
E. E. Larionova
Central Tuberculosis Research Institute
Russian Federation
Russian Federation
senior Researcher of Microbiological Department,
2, Yauzskaya alleya, Moscow, 107564
L. N. Chernousova
Central Tuberculosis Research Institute
Russian Federation
Russian Federation
leading Researcher, Head of Microbiological Department,
2, Yauzskaya alleya, Moscow, 107564
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Review
For citations:
Ustinova V.V., Smirnova T.G., Varlamov D.A., Andrievskaya I.Yu., Larionova E.E., Chernousova L.N. DETECTION AND DIFFERENTIATION OF NON-TUBERCULOUS MYCOBACTERIA AND M. TUBERCULOSIS COMPLEX BY REAL TIME PCR. Tuberculosis and Lung Diseases. 2016;94(9):80-87. (In Russ.) https://doi.org/10.21292/2075-1230-2016-94-9-80-87
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