STUDY DRUG SENSITIVITY M. TUBERCULOSIS WITH TEST SYSTEM SENSITITRE MYCOTB BASED ON CERTAIN CRITICAL CONCENTRATION FOR THE ANTITUBERCULOSIS DRUGS

Determination of the critical concentrations (CC) of drugs, widely used in the treatment of tuberculosis, to evaluate the results of the drug susceptibility testing of M. tuberculosis isolates using Sensititre MycoTB test system.
We studied using Sensititre MycoTB test system the minimal inhibitory concentrations (MICs) of streptomycin, isoniazid, rifampicin, ethambutol, amikacin, kanamycin, moxifloxacin and ofloxacin on the panel of the conditionally sensitive and conditionally resistant MBT strains isolated from different diagnostic material obtained in patients with pulmonary tuberculosis who were treated at the Moscow research and Clinical Center for Tuberculosis Control.
As a result of the analysis of the obtained MICs values, we determined the critical concentrations of antituberculosis drugs by MycoTB test system as the lowest concentration which inhibits the growth of 95% of sensitive MBT strains and allows the growth of 95% resistant MBT strains. We determined the following values of CCs: streptomycin – 1,0, isoniazid – 0,25, rifampicin – 1,0, ethambutol – 4,0, ofloxacin – 2,0, moxifloxacin – 0,25, kanamycin – 2,5 and amikacin – 1,0 μg/ml.
We carried out the comparative analysis of the effectiveness of the drug susceptibility testing (DST) of M.tuberculosis performed by Sencsititre MycoTB test in comparison with the cultivation methods on Middlebrook in the BACTEC™ MGIT™ 960 system and on Löwenstein-Jensen solid medium.
The cultures of M.tuberculosis obtained from 137 patients were investigated. There was the high percentage of matches of data obtained by Sencsititre MycoTB test and other methods. The Sencsititre MycoTB test has some advantages such as the standardization of the process and the ability of simultaneous quantitative determination of the drug resistance level for 12 antituberculosis drugs in the short period (7-14 days). The method of the DST by Sencsititre MycoTB test is simpler then others. This Sencsititre MycoTB test-system was tested abroad and in Moscow TB Control Center, certified in Russia and can be used in clinical practice.

1. Дорожкова И.Р. Лекарственная чувствительность микобактерий туберкулеза. Методы определения. Оценка результатов. / Лабораторные исследования при туберкулезе / под ред. В.И. Литвинова, А.М. Мороза. – М.: МНПЦБТ, 2013. – С. 80-101.

2. Исаева Ю.Д., Крылова Л.Ю., Букатина А.А. и др. Выявление Mycobacterium tuberculosis с широкой лекарственной чувствительностью. / Лабораторные исследования при туберкулезе / под ред. В.И. Литвинова, А.М. Мороза. – М.: МНПЦБТ, 2013. – С. 111–120.

3. О совершенствовании противотуберкулезных мероприятий в Российской Федерации: приказ Минздрава России от 21 марта 2003 г. № 109. Приложение № 11. Инструкция по унифицированным методам микробиологических исследований при выявлении, диагностике и лечении туберкулеза. [Электронный ресурс] URL: http://www.consultant.ru/document/cons_doc_LAW_100873/ (Дата обращения 25.04.2015).

4. Abuali M.M., Katariwala R., LaBombardi V.J. A comparison of the Sensititre® MYCOTB panel and the agar proportion method for the susceptibility testing of Mycobacterium tuberculosis // Eur. J. Clin. Microbiol. Infect. Dis. – 2011. – Vol. 31. – N. 5. – P. 835–839.

5. Ahuja S.D., Ashkin D., Avendano M. et al. Multidrug resistant pulmonary tuberculosis treatment regimens and patient outcomes: an individual patient data meta-analysis of 9,153 patients // PLoS Med. – 2012. – Vol. 9. – N. 8. – e1001300.

6. Banu S., Rahman S.M., Khan M.S. et al. Discordance across several methods for drug susceptibility testing of drug-resistant Mycobacterium tuberculosis isolates in a single laboratory // J. Clin. Microbiol. – 2014. – Vol. 52. – N. 1. – P. 156–63.

7. Global tuberculosis report 2013. – Geneva: WHO, 2013. [Электронный ресурс] URL: http://www.who.int/tb/publications/global_report/en/index.hyml. (Дата обращения 25.04.2015).

8. Guidelines for the programmatic management of drug-resistant tuberculosis: 2011 update. – Geneva: WHO, 2011.

9. Hall L., Jude K.P., Clark S.L. et al. Antimicrobial susceptibility testing of Mycobacterium tuberculosis complex for first and second line drugs by broth dilution in a microtitre plate format // J. Vis. Exp. – 2011. – Vol. 24. – N. 52. – P. 1–4.

10. Hall L., Jude K. P., Clark S.L. et al. Evaluation of the Sensititre MycoTB plate for susceptibility testing of the Mycobacterium tuberculosis сomplex against firstand second-line agents // J. Clin. Microbiol. – 2012. – Vol. 50. – N. 11. – P. 3732–3734.

11. Heifets L. Qualitative and quantitative drug-susceptibility tests in mycobacteriology // Am. Rev. Respir. Dis. – 1998. – Vol. 137. – P. 1217–1222.

12. Lee J., Armstrong D., Ssengooba W. et al. Sensititre MYCOTB MIC plate for testing Mycobacterium tuberculosis susceptibility to first- and second – line drugs // Antimicrob. Agents Chemother. – 2014. – Vol. 58. – N. 1. – P. 11–18.

13. Mpagama S., Houpt E., Stroup S. et al. Application of quantitative second-line drug susceptibility at a multidrug-resistant tuberculosis hospital in Tanzania // BMC Infectious Diseases. – 2013. – Vol. 13. – P. 432–441.

14. Siddiqi S., Rüsch-Gerdes S. MGIT Procedure Manual for BactecTM MGITTM 960 TB System (Also applicable for Manual MGIT) Mycobacteria Growht Indicator Tube (MGIT) / Culture and Drug Susceptibility Demonstration Projects. – Foundation for Innovative New Diagnostics, 2007. [Электронный ресурс] URL: http://www.ipaqt.org/wp-content/uploads/2013/02/MGIT-Procedure-Manual.pdf. (Дата обращения 25.04.2015).

15. Use of liquid TB culture and drug susceptibility testing (DST) in low and medium income settings. Summary report of the Expert Group Meeting on the use of liquid culture media, WHO Stop TB Department. – Geneva: WHO, 2007. [Электронный ресурс] URL: http://www.who.int/tb/dots/laboratory/Use of Liquid TB Culture_Summary Report.pdf. (Дата обращения 25.04.2015).