Phthisiatry
doi: 10.25005/2074-0581-2019-21-3-467-471
COMPLEX RESEARCH OF THE SPUTUM IN NEWLY DIAGNOSED PATIENTS WITH PULMONARY TUBERCULOSIS USING POLYMERASE CHAIN REACTION AND MYCROBIOLOGICAL CONTROL OF THE VIABILITY OF MYCOBACTERIUM TUBERCULOSIS
Laboratory of Immunology and Molecular Biology, National Center for Phthisiology, Bishkek, Kyrgyz Republic
Objective: The use of polymerase chain reaction (PCR) in a comprehensive study of sputum in newly diagnosed patients with pulmonary tuberculosis (PTB) in the control of mycobacterium tuberculosis (MTB) viability by the microbiological method.
Methods: The object of the study – 59 newly diagnosed patients with PTB in admission for treatment and 28 patients with PTB two months after the start of anti-tuberculosis chemotherapy. The study material is the sputum of PTB patients. Identification of the dead and persistent MTB was noted with the positive result of PCR (PCR+) and the absence of growth of MTB on the dense of nutritional environment of Levenstein-Jensen. The DNASorb-B and Litekh sets were used for DNA extraction of Mycobacterium tuberculosis complex. For amplification, Politub and AmpliSens MBT kits were used. Detection of amplification products was carried out with electrophoresis in 1.7% agarose gel in the presence of bromide ethidium
Results: Before treatment, PCR was detected in 52 (88.1%) of the 59 PTB patients examined, 34 (57.6%) tested positive for inoculation (culture+). 25 (42.4%) of patients, lack of growth of MTB out of 59 PTB patients. After 2 months of chemotherapy, PCR+ was detected in 23 (82.1%) out of 28 patients with PTB, culture+ – in 13 (46.4%), no growth of MTB was revealed in 15 (53.6%).
Conclusion: Before treatment, PCR was detected in 52 (88.1%) of the 59 PTB patients examined, 34 (57.6%) tested positive for inoculation (culture+). 25 (42.4%) of patients, lack of growth of MTB out of 59 PTB patients. After 2 months of chemotherapy, PCR+ was detected in 23 (82.1%) out of 28 patients with PTB, culture+ – in 13 (46.4%), no growth of MTB was revealed in 15 (53.6%).
Keywords: Pulmonary tuberculosis, polymerase chain reaction, microbiological method, mycobacterium tuberculosis viability, chemotherapy.
References
- Seryogina VA, Budritsky AM. Sovremennye vozmozhnosti diagnostiki tuberkulyoza lyogkikh [Modern possibilities of diagnosis of pulmonary tuberculosis]. Vestnik VGMU. 2016;15(4):7-17.
- Solovyova TN, Kozlova NV, Elkin AV, Barnaulov AO. Diagnostika infil’trativnogo tuberkulyoza lyogkikh v sovremennykh usloviyakh [Diagnosis of infiltrative pulmonary tuberculosis in modern conditions]. Vestnik SZGMU im. I.I. Mechnikova. 2013;5(3):79-83.
- Selivanov EV, Zvyagintsev EN. Mesto PTSR-issledovaniy v svoevremennoy i tochnoy diagnostike tuberkulyoza [Place of PCR studies in the timely and accurate diagnosis of tuberculosis]. Vestnik laboratorii DNK-diagnostiki. 2011;10:12-7.
- Costa P, Botelho A, Couto I, Viveiros M, Inacio J. Standing of nucleic acid testing strategies in veterinary diagnosis laboratories to uncover Mycobacterium tuberculosis complex members. Front Mol Biosci. 2014;1:1-18.
- Galimi R. Extrapulmonary tuberculosis: tuberculous meningitis new developments. Eur Rev Med Pharmaco. 2011;4:365-86.
- Sivokozov I, Shumskaya I, Lovacheva O, Evgushenko E, Chernousova L. Efficacy of endoscopic diagnostics in smear-negative TB. Eur Respire J. 2014;44:26-30.
- Ivanov PL, Leonov SL. Metodika polucheniya DNK, prigodnoy dlya molekulyarno-geneticheskogo issledovaniya, iz geparinizirovannykh obraztsov krovi [Method obtaining DNA suitable for molecular genetic research from heparinized blood samples]. Sudebno-meditsinskaya ekspertiza. 2010;6:21-3.
- Sevastyanova EV, Puzanov VA, Smirnova TG, Larionova EE, Chernousova LN. Otsenka kompleksa mikrobiologicheskikh i molekulyarno-geneticheskikh issledovaniy dlya diagnostiki tuberkulyoza [Assessment of a set of microbiological and molecular genetic studies for the diagnosis of tuberculosis]. Tuberkulyoz i bolezni lyogkikh. 2015;1:35-41.
- Sevastyanova EV, Larionova EE, Smirnova TG, Andrievskaya IU, Andreevskaya SN, Chernousova LN. Otsenka rezul’tatov vyyavlenia mikobakteriy, poluchennykh raznymi metodami issledovaniya [Evaluation of the result of Mycobacterium detection, obtained by different studies methods]. Meditsinskiy al’yans. 2018;3:25-30.
- Singh S, Saluja TP, Kaur M, Khilnani GC. Comparative evaluation of FASTP assay with PCR and other conventional in vitro diagnostic methods for the early detection of pulmonary tuberculosis. J Clin Lab Anal. 2008;22(5):367-74.
- Sevastyanova EV, Chernousova LN. Sovremennye algoritmy mikrobiologicheskoy diagnostiki tuberkulyoza [Modern algorithms of microbiological diagnostics of tuberculosis]. Tuberkulyoz i bolezni lyogkikh. 2018;96(7):11-7.
- Ekrami A, Samarbaf-Zadeh AR, Khosravi A, Zardar B, Alavi M, Amin M. Validity of bioconjugated silica nanoparticles in comparison with direct smear, culture and polymerase chain reaction for detection of mycobacterium tuberculosis in sputum specimens. Int J Nanomed. 2011;6:2729-35.
- Miotto P, Bigoni S, Migliori GB, Mattelli A, Cirillo DM. Early tuberculosis treatment monitoring by Xpert® MTB/RIF. Eur Respire J. 2012;39(5):1269-71.
- Velayat AA, Farnia P, Masiedi MR. Recurrence after treatment success in pulmonary multidrug resistant tuberculosis: predication by continual PCR positivity. Int J Clin Exp Med. 2012;5(3):271-2.
- Kondratyeva TC, Azhikina TL, Shleeva MO, Caprelyants AS, Apt AS. Geneticheskiy kontrol’ latentnoy tuberkulyoznoy infektsii [Genetic control of latent tuberculosis infection]. Tuberkulyoz i sotsial’no-znachimye zabolevaniya. 2013;2:61-7.
- Popov SA, Sabgayda TP, Mozhokina GN. Laboratornaya diagnostika bol’nykh tuberkulyozom s lekarstvennoy ustoychivost’yu vozbuditelya [The laboratory diagnostic of patients with tuberculosis with medicinal resistance of agent]. Problemy sotsial’noy gigieny, zdravookhraneniya i istorii meditsiny. 2016;24(5):272-6.
- Khomenko DS, Tigiev AV, Campos ED. Epidemiologicheskie i kliniko-rentgenologicheskie osobennosti infil’trativnogo tuberkulyoza lyogkikh v Rostovskoy oblasti [Epidemiological and clinicoroentgenological features of infiltrative pulmonary tuberculosis in Rostov region]. Zhurnal fundamental’noy meditsiny i biologii. 2014;3:49-51.
- Savintseva EV, Yakovleva EA, Shevelyova SL, Grinko OV, Svetlakova AA. Retrospektivnyy analiz metodov diagnostiki – BACTEC, klassicheskiy metod, PTSRmetod [Retrospective analysis of methods of diagnostics – BACTEC, classical method, PCR-method]. European Science. 2019;2:46-9.
Authors' information:
Dudenko Elena Vyacheslavovna
Senior Researcher, Laboratory of Immunology and Molecular Biology, National Center for Phthisiology
ORCID ID: 0000-0001-8948-3659
Sydykova Saltanat
Researcher, Laboratory of Immunology and Molecular Biology, National Center for Phthisiology
ORCID ID: 0000-0001-6479-2770
Information about support in the form of grants, equipment, medications
The work was carried out according to the plan of scientific research works of National Center for Phthisiology (state registration number – 0003059). The authors did not receive financial support from manufacturers of medicines and medical equipment.
Conflicts of interest: No conflict
Address for correspondence:
Dudenko Elena Vyacheslavovna
Senior Researcher, Laboratory of Immunology and Molecular Biology, National Center for Phthisiology
720020, Kyrgyz Republic, Bishkek, Akhunbaev Street, 90a
Tel.: +996 (554) 044011
E-mail: dudenko-1963@mail.ru