Using a new molecular genetic of genotype and liquid culture medium for rapid diagnosis tb
DOI:
https://doi.org/10.15587/2313-8416.2015.51771Keywords:
tuberculosis, model, treatment, isoniazid, dose, experiment, change, system, research, diagnosticAbstract
This paper presents the results of molecular genetic test system GenoType multyresistentens MTBDRplus. It was established that the presence of mutations associated with resistance to isoniazid, only 93.1 % of cases of MBT to isoniazid during the test in a liquid medium. Work carried out under the National Programme to combat tuberculosis
Materials and methods. We investigated the clinical sputum samples from patients with pulmonary tuberculosis. The applied system GenoType. Principle DNA strip technology GenoType is that the DNA-coated strip specific test that are complementary to the derived PCR amplicon. After the single-stranded amplicon denaturation associated with tests on strip (hybridize), and visualized in a sequential enzymatic reaction with streptavydynom and alkaline phosphatase. Evaluation of hybridization is performed automatically. For culturing sputum liquid culture medium used - Middlebrook broth 7N9 VASTES MGIT system.
Results and discussion. The results of molecular genetic studies of samples of sputum-concentrated and concentrated by a system GenoType not differed (P>0.05). Diagnostic value of two methods (molecular and genetic – system GenoType and phenotype – VASTES MGIT 960 system) was very high (100%). Two systems have tested positive in the study 756 (95.5 %) Mycobacterium strains that were identified in the system VASTES MGIT 960, formed Cord Factor and the results were positive identification test ID MTB MGIT they attributed to Mycobacterium tuberculosis complex. 36 (4.5 %) samples from positive MGIT tubes were negative. As a result of molecular-genetic identification of nontuberculous mycobacteria complex it was found that 18 (2.3 %) strains of mycobacteria belonging to the M. avium-intracellulare, 12 (1.5 %) mycobacterial cultures were attributed to M. kansasii, 6 (0, 7 %) cultures were identified as M. fortuitum. The results of the molecular study of MS on Mycobacterium resistance profile INN + RIF coincided in 95.5 % (894 strains) the results of testing by phenotypic proportions. In the presence of mutations associated with resistance to INH, only 93.1 % of cases observed MC M. tuberculosis to INH during TMCH in a liquid medium Middlebrook 7N9. In the presence of mutations in the genes responsible for resistance to the presence of Q, in a liquid medium only 288 (90.6 %) strains of M. tuberculosis have MS to Ofx. The strains of mycobacteria DNA were detected mutations in genes associated with MS to aminoglycosides / cyclic peptides in 299 (94.0%) cases were resistant to Am and 302 (94.9 %) cases were resistant to the results Cm DST system VASTES MGIT 960. In determining the resistance to E major differences were found between the productivity of molecular genetic and phenotypic research methods – only 206 (64, 2 %) strains of M. tuberculosis were resistant to the analysis system VASTES MGIT 960.
Conclusions. GenoType system allows you to quickly carry out the identification and differentiation of Mycobacterium strains of nontuberculous mycobacteria. The results of the molecular genetic studies multyrezystents system GenoType MTBDRplus match in 95,5 % of the phenotypic test results by proportions. Using DNA strip technology GenoType to determine mutations in the genes of Mycobacterium responsible for the IPU to TDC, necessarily requires a parallel DST setting in a liquid medium
References
Yrtuhanova, A. A. et. al (2001). Automated methods for determining Mousse culture Bacterium tuberculosis in zhydkyh environments. Problems of tuberculosis, 3, 53–56.
Dorozhkova, R. I., Makarov, M. V., Freiman, G. E. (2012). Allocation Increase of the effectiveness and Authentication mykobakteryy in terms tsentralyzovannoy mykobakteryolohycheskoy laboratory. Tuberculosis and lung disease, 11, 21–26.
Barbova, A. I. et. al (2007). The use of automated MGIT system for the diagnosis of pulmonary tuberculosis and determination of drug resistance of mycobacteria. Institute of Phthisiology and Pulmonology. F. G. Yanovsky AMS Ukraine. Kyiv: IFS, 24.
Zhurylo, O. A., Barbova, A. I., Trofimova, P. S. et. al (2013). Molecular genetic approaches to identify mycobacteria.V Congress of TB doctors and pulmonologists Ukraine. Kyiv, 123–124.
Eliseev, P. I., Nikishova, E. I., Gorina, G. P., Mar'jandyshev, A. O. (2012). Results of application methods GenoType MTBDRplus and BASTES MGIT lekarstvennoy chuvstvytelnosty definitions for tuberculosis pathogen. Tuberculosis and lung disease, 6, 31–34.
Zhurylo, A. A. (2013). Quality Assurance System bacteriological research in institutions that carry out microbiological diagnosis of tuberculosis at different levels of care Ukraine. Kirovograd: Polium, 72.
Zhurilo, O. A., Barbova, A. І., Glushkevich, T. G., Tretjakova, L. V. (2012). Standards bacteriological diagnosis of tuberculosis in laboratories TB facilities Ukraine. Kirovograd: Polium, 190.
Barbova, A. I., Zhurylo, O. A., Trofimova, P. S. et. al (2013). Modern approaches to conducting bacteriological identification of Mycobacterium. Ukr. pulmonol. Zh., 3, 28–32.
Feschenko, Y. I., Cherenko, A., Barbova, A. I. (2013). Unified clinical protocols of primary, secondary (specialized) and tertiary (highly specialized) medical care "Tuberculosis", especially its training and different from previous clinical protocols. Tuberculosis, pulmonary disease, HIV infection, 2, 8–18.
Kulikov, N. V., Kaplina, M. L., Kalinin, O. L. (1996). Preparation and characterization of antibodies to Mycobacterium monoklonovyh Stegmatis and Kansassi. Problems tuberkuleza, 1, 2–4.
Cherenko, S. O. (2010). Basic principles of treatment of tuberculosis resistant Mycobacterium tuberculosis. Ukr. Pul monological magazine, 3, 27–32.
Lu, P.-L., Yang, Y.-C., Huang, S. C., Jenh, Y.-S., Lin, Y.-C., Huang, H.-H., Chang, T. C. (2011). Evaluation of the Bactec MGIT 960 System in Combination with the MGIT TBc Identification Test for Detection of Mycobacterium tuberculosis Complex in Respiratory Specimens: Table 1. Journal of Clinical Microbiology, 49 (6), 2290–2292. doi: 10.1128/jcm.00571-11
Rusch-Gerdes, S., Pfyffer, G. E., Casal, M., Chadwick, M., Siddiqi, S. (2006). Multicenter Laboratory Validation of the BACTEC MGIT 960 Technique for Testing Susceptibilities of Mycobacterium tuberculosis to Classical Second-Line Drugs and Newer Antimicrobials. Journal of Clinical Microbiology, 44 (3), 688–692. doi: 10.1128/jcm.44.3.688-692.2006
Kiet, V. S., Lan, N. T. N., An, D. D., Dung, N. H., Hoa, D. V., van Vinh Chau, N. et. al (2010). Evaluation of the MTBDRsl Test for Detection of Second-Line-Drug Resistance in Mycobacterium tuberculosis. Journal of Clinical Microbiology, 48 (8), 2934–2939. doi: 10.1128/jcm.00201-10
Lacoma, A., García-Sierra, N., Prat, C., Maldonado, J., Ruiz-Manzano, J., Haba, L. Et. al (2011). GenoType MTBDR sl for Molecular Detection of Second-Line-Drug and Ethambutol Resistance in Mycobacterium tuberculosis Strains and Clinical Samples. Journal of Clinical Microbiology, 50 (1), 30–36. doi: 10.1128/jcm.05274-11
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Copyright (c) 2015 Людмила Володимирівна Гайова, Ганна Іванівна Барбова, Олександр Анатолійович Журило, Поліна Станіславівна Трофимова, Наталія Миколаївна Алієва
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