Identification of rpoB Gene Mutations in MDR Mycobacterium tuberculosis Isolates Using Real-time PCR in Semarang
Published 2026-10-01
Keywords
- Mycobacterium Tuberculosis,
- rpoB gene,
- rifampicin resistance,
- MDR-TB ,
- Real-time PCR
How to Cite
Copyright (c) 2026 Jaringan Laboratorium Medis

This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Abstract
Indonesia is among the countries with a high burden of multidrug-resistant tuberculosis (MDR-TB). The majority of rifampicin resistance in Mycobacterium tuberculosis (>95%) is caused by mutations in the rpoB gene which encodes the beta subunit of RNA polymerase specifically within an 81-base-pair "hotspot" region. This study aimed to identify rpoB gene mutations in MDR-TB isolates using real-time PCR. It was a descriptive observational study employing a laboratory-based approach. Samples were selected using a consecutive sampling technique, consisting of 40 sputum specimens from TB-positive patients. Sample collection was conducted at the Community Health Centers (Balkesmas) in the Semarang City and Ambarawa regions, while mutation analysis was performed at the Laboratory of the Medical Laboratory Technology Department, Poltekkes Kemenkes Semarang. The results showed that the rpoB gene drug dominated as a marker of rifampicin resistance detected in 87.5% of samples (35 out of 40). Five samples yielded no detection results, likely due to target DNA degradation during the study process. This study demonstrates the presence of rpoB gene mutations as an indicator of rifampicin resistance in TB cases within the Semarang and Ambarawa regions.
References
- Adikaram CP, Perera J, Wijesundera SS. Geographical profile of rpoB gene mutations in rifampicin resistant Mycobacterium tuberculosis isolates in Sri Lanka. Microb Drug Resist. (2012) Oct;18(5):525-30. doi: 10.1089/mdr.2012.0031. Epub 2012 Jun 25. PMID: 22731859.
- Amalia, E., Nindatama, M. R., Hayati, L., & Handayani, D. (2015). Identifikasi Mutasi Gen rpob Ser531Leu Mycobacterium tuberculosis Yang Berhubungan Dengan Resistensi Rifampisin. Biomedical Journal of Indonesia, 1(1), 30-34.
- Bostanabad SZ, Fateh A, Seyedi K, Abdolrahimi F, Karimi, Tasbiti AH, et al. (2007) Frequency and molecular characterization of rifampicin-resistance in rpoB region of Multiple Drug Resitance (MDR) isolates from tuberculosis patients in southern endemic region of Iran. Iranian Journalof Biotechnology 5(4):212-217.
- Brooks GF, Butel JS, Morse SA. Mikrobiologi Kedokteran Jawetz, Melnick, and Adelberg (Ed 23rd). (2008). Terjemah oleh: Hartanto, H, et al. EGC, Jakarta, Indonesia, Hal: 325-333.
- Comas I, Borrell S, Roetzer A, Rose G, Malla B, Maeda MK, et al. (2012). Whole-Genome Sequencing of Rifampicin-Resistant M. Tuberculosis Strains Identifies Compensatory Mutations In RNA Polymerase. Nat Genet; 44(1):106–110.
- Isakova, J., Sovkhozova, N., Vinnikov, D., Goncharova, Z., Talaibekova, E., Aldasheva, N., & Aldashev, A. (2018). Mutations of rpoB, katG, inhA and ahp genes in rifampicin and isoniazid-resistant Mycobacterium tuberculosis in Kyrgyz Republic. BMC microbiology, 18(1), 22.
- Jiao W-W, Mokrousov I, Sun G-Z, Li M, Liu J-W, Narvskaya O, et al. (2007). Molecular characteristics of rifampin and isoniazid resistant Mycobacterium tuberculosis strains from Beijing. China Chin Med J (Engl). 120:814–9.
- Jo K-W, Lee S, Kang MR, Sung H, Kim M-N, Shim TS. (2017). Frequency and type of disputed rpoB mutations in Mycobacterium tuberculosis isolates from South Korea. Tuberc Respir Dis. 80:270–6.
- Kementerian Kesehatan RI. (2023). Pertemuan Monitoring dan Evaluasi Program Tuberkulosis Tahun 2023. DKI Jakarta.
- Kementerian Kesehatan RI. (2023). Laporan Program Penanggulangan Tuberkulosis Tahun 2022. Program Penanggulangan Tuberkulosis. Jakarta :Kementerian Kesehatan RI. 2023.
- Minh NN, Van Bac N, Son NT, Lien VTK, Ha CH, Cuong NH, et al. (2012). Molecular characteristics of rifampin-and isoniazid-resistant Mycobacterium tuberculosis strains isolated in Vietnam. J Clin Microbiol. 50:598–601.
- Paluch-Oles, Jolanta, Maria Kozioł-Montewka, and Agnieszka Magrys. (2009). "Mutations in the rpoB gene of rifampin-resistant Mycobacterium tuberculosis isolates from Eastern Poland." The new microbiologica 32, no. 2 147-152.
- Ramaswamy S, Musser JM. (1998). Molecular genetic basis of antimicrobial agent resistance in Mycobacterium tuberculosis: 1998 update. Tuber Lung Dis. 79(1):3-29.
- Rosales-Klintz S, Jureen P, Zalutskayae A, Skrahina A, Xu B, Hu Y, et al. (2012). Drug resistance-related mutations in multidrug-resistant Mycobacterium tuberculosis isolates from diverse geographical regions. Int J Mycobacteriology. 1:124–30
- Rosilawati, M. L. (2007). Deteksi Mycobacterium tuberculosis dan resistensinya terhadap rifampisin dengan Metode nested polymerase chain reaction (PCR) dan sequencing. Universa Medicina, 26(1), 1-10.
- Sajduda A, Brzostek A, Popławska M, Augustynowicz-Kopeć E, Zwolska Z, Niemann S, et al. (2004). Molecular characterization of rifampin-and isoniazid-resistant Mycobacterium tuberculosis strains isolated in Poland. J Clin Microbiol. 42:2425–31.
- Syaifudin M, Rosilawati M, Irawan H, Bela B. (2007). Identifikasi Mycobacterium tuberculosis dan analisis mutasi gen rpoB dan katG penyebab resistensi ganda dengan teknik molekuler. Laporan Penelitian Balitbang BATAN. Jakarta, 1-15.
- Tavanaee Sani A, Ashna H, Kaffash A, Khaledi A, Ghazvini K. (2018). Mutations of rpob Gene Associated with Rifampin Resistance among Mycobacterium Tuberculosis Isolated in Tuberculosis Regional Reference Laboratory in Northeast of Iran during 2015-2016. Ethiop J Health Sci. May;28(3):299-304. doi: 10.4314/ejhs.v28i3.7. PMID: 29983529; PMCID: PMC6016358.
- Telenti, A., Imboden, P., Marchesi, F., Matter, L., Schopfer, K., Bodmer, T., ... & Cole, S. (1993). Detection of rifampicin-resistance mutations in Mycobacterium tuberculosis. The Lancet, 341(8846), 647-651.
- World Health Organization. (2022). Global Tuberculosis Report 2022. Geneva: World Health Organization.
- WHO. (2023). Global Tuberculosis Report 2023.
- Yue, J., Shi, W., Xie, J., Li, Y., Zeng, E., & Wang, H. (2003). Mutations in the rpoB gene of multidrug-resistant Mycobacterium tuberculosis isolates from China. Journal of clinical microbiology, 41(5), 2209-2212
- Yuwono T. (2005). Biologi Molekuler. Erlangga, Jakarta, Indonesia.
- Zaw MT, Emran NA, Lin Z. (2018). Mutations inside rifampicin-resistance determining region of rpoB gene associated with rifampicin-resistance in Mycobacterium tuberculosis. J Infect Public Health. 2018 Sep-Oct;11(5):605-610. doi: 10.1016/j.jiph.2018.04.005. Epub 2018 Apr 26. PMID: 29706316.
