Molecular epidemiological typing of M. tuberculosis isolates isolated from Turkey's Eastern Anatolia with in house PCR method
Öz
Purpose: The aim of this study as to compare microscopy and culture results of samples, determine drug resistance rates of the isolates, evaluate epidemiological relationship between the strains with ERIC-PCR, RAPD-PCR, OUT-PCR based on in house PCR technique.
Materials and Methods: Direct microscopy and culture results of 2010 samples were analyzed. Drug sensitivity results were obtained from TULSA. The typing of isolates based on in house PCR was carried out in the microbiology laboratory of a faculty of medicine in a state university.
Results: Of positive samples, 2.68% had Acid-resistant bacilli (ARB) positive + culture positivity, 2.93% had ARB positive and culture negative in smear, and 1.34% had ARB negative and culture positivity in smear. Resistance to primary antituberculous (anti-TB) drugs wasn’t observed in 33 culture positive isolates, whereas resistance to one or more primary anti-TB drugs was observed in 9.09%. Single drug resistance was 3.03%, resistance to Isoniazid (INH) and INH critical drugs was 6.06%. Isolates were divided 3 groups by ERIC-PCR, 5 groups for OUT-PCR and 6 by RAPD-PCR.
Conclusion: Positivity rates were low due to low rate of studied samples, negative samples taken during treatment process. Molecular techniques like ERIC-PCR, RAPD-PCR and OUT-PCR are easy, fast and inexpensive methods for the epidemiological typing of Mycobacterium tuberculosis (MTB) in evaluating distinctions, similarities between origins.
Anahtar Kelimeler
Mycobacterium tuberculosis, EZN, ERIC-PCR, RAPD-PCR, OUT-PCR
Kaynakça
- World Health Organization. Global Tuberculosis Report, 2020. Geneva, WHO, 2020.
- T.C. Ministry of Health General Directorate of Public Health Tuberculosis Department. TB War in Turkey 2018 Report. Ankara, Ministry of Health, 2018.
- T.C. Ministry of Health General Directorate of Public Health Tuberculosis Department, TB war in Turkey 2019 Report. Ankara, Ministry of Health, 2020.
- Dheda K, Ruhwald M, Theron G, Peter J, Yam WC. Point-of-care diagnosis of tuberculosis: past, present and future. Respirology. 2013;18:217-32.
- Perkins MD, Cunningham J. Facing the crisis: improving the diagnosis of tuberculosis in the HIV era. J Infect Dis. 2007;15:15-27.
- Cattamanchi A, Davis JL, Pai M, Huang L, Hopewell PC, Steingart KR. Does bleach processing increase the accuracy of sputum smear microscopy for diagnosing pulmonary tuberculosis? J Clin Microbiol. 2010;48:2433-9.
- Alp A. Current status in laboratory diagnosis of tuberculosis. Hacettepe Tıp Dergisi. 2011;42:28-33.
- Meehan CJ, Goig GA, Kohl TA, Verboven L, Dippenaar A, Ezewudo M et al. Whole genome sequencing of Mycobacterium tuberculosis: current standards and open issues. Nat Rev Microbiol. 2019;17:533-45.
- Igarashi Y, Chikamatsu K, Aono A, Yi L, Yamada H, Takaki A et al. Laboratory evaluation of the Anyplex II MTB/MDR and MTB/XDR tests based on multiplex real-time PCR and melting-temperature analysis to identify Mycobacterium tuberculosis and drug resistance. Diagn Microbiol Infect Dis. 2017;89:276-81.
- Ruiz P, Causse M, Vaquero M, Gutierrez JB, Casal M. Evaluation of a new automated Abbott RealTime MTB RIF/INH assay for qualitative detection of rifampicin/isoniazid resistance in pulmonary and extra-pulmonary clinical samples of Mycobacterium tuberculosis. Infect Drug Resist. 2017;6:463-7.
