Mycobacteriophage-based test system for phenotypic drug sensitivity of clinical isolates of tuberculous mycobacteria
The issue of rapid phenotypic drug sensitivity testing of clinical isolates of tuberculous mycobacteria remains relevant.The objective of the study is to develop a new test system for rapid phenotypic drug sensitivity testing of MTB clinical isolates based on lytic mycobacteriophages, capable of tes...
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| Format: | Article |
| Language: | Russian |
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New Terra Publishing House
2020-09-01
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| Series: | Туберкулез и болезни лёгких |
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| Online Access: | https://www.tibl-journal.com/jour/article/view/1448 |
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| author | M. B. Lapenkova G. A. Аrustamova Yu. S. Аlyapkina P. N. Filippov S. V. Lazebny M. A. Vladimirskiy |
| author_facet | M. B. Lapenkova G. A. Аrustamova Yu. S. Аlyapkina P. N. Filippov S. V. Lazebny M. A. Vladimirskiy |
| author_sort | M. B. Lapenkova |
| collection | DOAJ |
| description | The issue of rapid phenotypic drug sensitivity testing of clinical isolates of tuberculous mycobacteria remains relevant.The objective of the study is to develop a new test system for rapid phenotypic drug sensitivity testing of MTB clinical isolates based on lytic mycobacteriophages, capable of testing resistance to first and second line drugs.Subjects and methods. Cultures of MTB (108 clinical isolates after primary culturing in Bactec MGIT) were incubated for 48 hours with addition of anti-tuberculosis drugs, then for 48 hours more after adding lytic mycobateriophage D29. The subsequent multiplex reaction of the polymerase chain reaction in real time allowed performing quantitative analysis of MTB DNA and mycobacteriophage DNA. The drug sensitivity of the tested sample was assessed by ranges of the differences in fluorescence threshold levels corresponding to the amount of mycobacteriophages between the control and test sample. At the same time, the drug sensitivity/resistance of all MTB clinical isolates after repeated culture was tested by Bactec MGIT which was adopted as a reference method.Results. Lytic mycobacteriophage-based drug sensitivity testing of 108 MTB isolated to four first line drugs and 90 isolates to six second line anti-tuberculosis drugs demonstrated a high level of concordance with the results of the Bactec MGIT system. It provided 99.5% sensitivity and 100% specificity of the method for 3 first-line drugs; it was slightly lower for ethambutol (86 and 96.9%, respectively), while for second line drugs, its sensitivity made 94.83% and specificity - 98.85%. |
| format | Article |
| id | doaj-art-e70a38775479462ab922ee5c10d8c194 |
| institution | OA Journals |
| issn | 2075-1230 2542-1506 |
| language | Russian |
| publishDate | 2020-09-01 |
| publisher | New Terra Publishing House |
| record_format | Article |
| series | Туберкулез и болезни лёгких |
| spelling | doaj-art-e70a38775479462ab922ee5c10d8c1942025-08-20T01:47:52ZrusNew Terra Publishing HouseТуберкулез и болезни лёгких2075-12302542-15062020-09-01988142210.21292/2075-1230-2020-98-8-14-221447Mycobacteriophage-based test system for phenotypic drug sensitivity of clinical isolates of tuberculous mycobacteriaM. B. Lapenkova0G. A. Аrustamova1Yu. S. Аlyapkina2P. N. Filippov3S. V. Lazebny4M. A. Vladimirskiy5National Medical Research Center of Phthisiopulmonology and Infectious DiseasesNational Medical Research Center of Phthisiopulmonology and Infectious DiseasesNational Medical Research Center of Phthisiopulmonology and Infectious DiseasesMoscow Regional Clinical TB DispensaryMoscow Regional Clinical TB DispensaryNational Medical Research Center of Phthisiopulmonology and Infectious DiseasesThe issue of rapid phenotypic drug sensitivity testing of clinical isolates of tuberculous mycobacteria remains relevant.The objective of the study is to develop a new test system for rapid phenotypic drug sensitivity testing of MTB clinical isolates based on lytic mycobacteriophages, capable of testing resistance to first and second line drugs.Subjects and methods. Cultures of MTB (108 clinical isolates after primary culturing in Bactec MGIT) were incubated for 48 hours with addition of anti-tuberculosis drugs, then for 48 hours more after adding lytic mycobateriophage D29. The subsequent multiplex reaction of the polymerase chain reaction in real time allowed performing quantitative analysis of MTB DNA and mycobacteriophage DNA. The drug sensitivity of the tested sample was assessed by ranges of the differences in fluorescence threshold levels corresponding to the amount of mycobacteriophages between the control and test sample. At the same time, the drug sensitivity/resistance of all MTB clinical isolates after repeated culture was tested by Bactec MGIT which was adopted as a reference method.Results. Lytic mycobacteriophage-based drug sensitivity testing of 108 MTB isolated to four first line drugs and 90 isolates to six second line anti-tuberculosis drugs demonstrated a high level of concordance with the results of the Bactec MGIT system. It provided 99.5% sensitivity and 100% specificity of the method for 3 first-line drugs; it was slightly lower for ethambutol (86 and 96.9%, respectively), while for second line drugs, its sensitivity made 94.83% and specificity - 98.85%.https://www.tibl-journal.com/jour/article/view/1448tuberculous mycobacteriadrug sensitivitymycobacteriophagepolymerase chain reaction |
| spellingShingle | M. B. Lapenkova G. A. Аrustamova Yu. S. Аlyapkina P. N. Filippov S. V. Lazebny M. A. Vladimirskiy Mycobacteriophage-based test system for phenotypic drug sensitivity of clinical isolates of tuberculous mycobacteria Туберкулез и болезни лёгких tuberculous mycobacteria drug sensitivity mycobacteriophage polymerase chain reaction |
| title | Mycobacteriophage-based test system for phenotypic drug sensitivity of clinical isolates of tuberculous mycobacteria |
| title_full | Mycobacteriophage-based test system for phenotypic drug sensitivity of clinical isolates of tuberculous mycobacteria |
| title_fullStr | Mycobacteriophage-based test system for phenotypic drug sensitivity of clinical isolates of tuberculous mycobacteria |
| title_full_unstemmed | Mycobacteriophage-based test system for phenotypic drug sensitivity of clinical isolates of tuberculous mycobacteria |
| title_short | Mycobacteriophage-based test system for phenotypic drug sensitivity of clinical isolates of tuberculous mycobacteria |
| title_sort | mycobacteriophage based test system for phenotypic drug sensitivity of clinical isolates of tuberculous mycobacteria |
| topic | tuberculous mycobacteria drug sensitivity mycobacteriophage polymerase chain reaction |
| url | https://www.tibl-journal.com/jour/article/view/1448 |
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