The aim of the project: The aim of the project is to conduct a comprehensive molecular genetic characterization of drug resistance to pyrazinamide (PZA), the first-line anti-tuberculosis (anti-TB) drug in Kazakhstan
Project Description: According to WHO recommendations, first-line anti-TB drugs such as isoniazid, rifampicin, ethambutol and pyrazinamide are recommended for the treatment of drug-susceptible tuberculosis. All of these anti-TB drugs are widely used in the treatment of tuberculosis, but drug susceptibility testing for pyrazinamide (PZA) unlike the other three anti-TB drugs is not routinely performed due to problems associated with phenotypic drug susceptibility testing methods. In recent WHO recommendations, the non-radiometric BACTEC MGIT-960 test in combination with genetic testing need to be used to determine pyrazinamide resistance. Pyrazinamide also forms a key component of anti-TB drugs recommended for the treatment of MDR-TB. When combined with novel anti-TB drugs, PZA has good action due to the potential synergistic effect. That is why it is important to understand the PZA resistance patterns of M. tuberculosis.The real PZA resistance of M. tuberculosis circulating in Kazakhstan, the percentage of false resistance results, and possible interactions of phenotypic resistance with mutations in pncA and other rpsA and panD genes are unknown. There are no accurate data for PZA drug resistance in Kazakhstan and the generation of local knowledge is important for TB control. The project will generate new information and knowledge, which can change testing methods of diagnostic laboratories for PZA resistance and possibly lead to new approaches of TB treatment in Kazakhstan. It will also generate new knowledge regarding PZA resistance and improved methods of detection of various forms and genetic mechanisms that are involved in the formation of PZA resistance in Kazakhstan. Therefore, understanding drug resistance mechanisms is essential to improving the diagnosis and treatment of TB patients. It is quite important to generate and analyze local knowledge on drug resistance and molecular genotyping. Drug resistance surveillance is necessary and should be of the highest priority from the National TB Program.
PI: Ulan Kozhamkulov
Co-PI: Ainur Akhmetova, Dauren Yerezhepov, Zhannur Abilova
Co-PI: Ainur Akhmetova, Dauren Yerezhepov, Zhannur Abilova
Realisation Period: 2024-2026
Expected results: The project results will include:
- Drug-resistant clinical isolates of M. tuberculosis will be collected and phenotypic drug susceptibility will be determined;
- Genotyping will be conducted and biological diversity of collected drug-resistant clinical isolates of M. tuberculosis from different regions of Kazakhstan will be assessed based on genotyping (MIRU-VNTR analysis/spoligotyping);
- Distribution frequency of drug resistance of M. tuberculosis to pyrazinamide will be identified among drug-resistant clinical isolates, including MDR/XDR in Kazakhstan;
- The results of phenotypic and genotypic methods of determination of drug resistance of M.tuberculosis to pyrazinamide will be compared. Cases of inconsistency in pyrazinamide resistance results will be determined;
- Mutations in pncA gene of pyrazinamide-resistant clinical isolates of M. tuberculosis will be detected and possible mutations in rpsA and panD genes (alternative mechanisms) associated with resistance to PZA will be identified.
Scientific publications will be published:
- at least 3 (three) articles and (or) reviews in peer-reviewed scientific publications in the scientific direction of the project, indexed in the Science Citation Index Expanded and included in the 1 (first), 2 (second) and (or) 3 (third) quartile according to impact factor in the Web of Science database and (or) having a CiteScore percentile in the Scopus database of at least 60 (sixty);
- at least 1 (one) article or review in a peer-reviewed foreign or domestic publication recommended by KOKNVO;
- or at least 2 (two) articles and (or) reviews in peer-reviewed scientific publications indexed in the Science Citation Index Expanded and included in the 1st (first) and (or) 2nd (second) quartile by impact factor in the Web of Science database and (or) having a CiteScore percentile in the Scopus database of at least 70 (seventy);
- or at least 1 (one) article or review in a peer-reviewed scientific publication, indexed in the Science Citation Index Expanded and included in the 1st (first) quartile in the Web of Science database or having a CiteScore percentile in the Scopus database of at least 90 (ninety).
- Drug-resistant clinical isolates of M. tuberculosis will be collected and phenotypic drug susceptibility will be determined;
- Genotyping will be conducted and biological diversity of collected drug-resistant clinical isolates of M. tuberculosis from different regions of Kazakhstan will be assessed based on genotyping (MIRU-VNTR analysis/spoligotyping);
- Distribution frequency of drug resistance of M. tuberculosis to pyrazinamide will be identified among drug-resistant clinical isolates, including MDR/XDR in Kazakhstan;
- The results of phenotypic and genotypic methods of determination of drug resistance of M.tuberculosis to pyrazinamide will be compared. Cases of inconsistency in pyrazinamide resistance results will be determined;
- Mutations in pncA gene of pyrazinamide-resistant clinical isolates of M. tuberculosis will be detected and possible mutations in rpsA and panD genes (alternative mechanisms) associated with resistance to PZA will be identified.
Scientific publications will be published:
- at least 3 (three) articles and (or) reviews in peer-reviewed scientific publications in the scientific direction of the project, indexed in the Science Citation Index Expanded and included in the 1 (first), 2 (second) and (or) 3 (third) quartile according to impact factor in the Web of Science database and (or) having a CiteScore percentile in the Scopus database of at least 60 (sixty);
- at least 1 (one) article or review in a peer-reviewed foreign or domestic publication recommended by KOKNVO;
- or at least 2 (two) articles and (or) reviews in peer-reviewed scientific publications indexed in the Science Citation Index Expanded and included in the 1st (first) and (or) 2nd (second) quartile by impact factor in the Web of Science database and (or) having a CiteScore percentile in the Scopus database of at least 70 (seventy);
- or at least 1 (one) article or review in a peer-reviewed scientific publication, indexed in the Science Citation Index Expanded and included in the 1st (first) quartile in the Web of Science database or having a CiteScore percentile in the Scopus database of at least 90 (ninety).
Methodology: In this project, we are working with National Reference laboratory of the National Scientific Center for Phthisiopulmonology (NRL NSCP) of the Republic of Kazakhstan (former National TB Center in Almaty, Kazakhstan). NRL NSCP will collect clinical isolates from two biggest cities of Kazakhstan (Astana, Almaty) and from some regions of Kazakhstan (TB hospitals where carry out susceptibility to first line anti-TB drugs and susceptibility to PZA by using BACTEC MGIT 960 system). The Laboratory of NSCP will re-check PZA susceptibility of collected clinical isolates by using BACTEC MGIT-960 system, will extract DNA from clinical isolates of Mycobacterium tuberculosis and carry out standardization of inoculum size of PZA drug susceptibility testing on BACTEC MGIT 960 system.
The genetic research (WGS/DNA sequencing of mutations in pncA gene and mutations in genes RpsA, PanD lacking pncA mutations, genotyping by MIRU-VNTR analysis/spoligotyping) will be conducted at the Laboratory of Genomic and Personalized Medicine, National Laboratory Astana, Nazarbayev University.
The genetic research (WGS/DNA sequencing of mutations in pncA gene and mutations in genes RpsA, PanD lacking pncA mutations, genotyping by MIRU-VNTR analysis/spoligotyping) will be conducted at the Laboratory of Genomic and Personalized Medicine, National Laboratory Astana, Nazarbayev University.
Contacts: ulan.kozhamkulov@nu.edu.kz