Assessment of the Genexpert MTB/RIF Assay's Performance for the Quick Identification of Rifampicin-Resistant Mycobacterium Tuberculosis
Keywords:
GeneXpert MTB/RIF assay, Mycobacterium Tuberculosis, bacillary load, rifampicin resistance TB (RR-TB)Abstract
Introduction: Tuberculosis (TB) remains a global health crisis, and the emergence of rifampicin-resistant TB (RR-TB) has further escalated this challenge. The speed and accuracy of diagnosis are critical for the timely treatment of the disease. Objective: The purpose of this study was to evaluate the diagnostic performance of the GeneXpert MTB/RIF assay in identifying MTB and rifampicin resistance, in an effort to enhance the diagnosis and treatment of tuberculosis (TB).Materials and Methods: This cross-sectional study was conducted at the National Tuberculosis Institute (NTI)/National Reference Laboratory (NRL) for Tuberculosis in Baghdad from February to September 2021. Morning sputum samples were collected from consenting patients. These samples were analyzed using Ziehl-Neelsen staining microscopy, the Xpert MTB/RIF assay, and culture on Löwenstein-Jensen (LJ) medium. Results: Among 140 suspected TB patients, 60 were confirmed to have active pulmonary tuberculosis (ATB) based on at least one positive microbiological test result, including acid-fast bacilli (AFB) staining, bacterial culture, molecular testing (Xpert MTB/RIF), clinical symptoms, and chest radiography (CXR) indicative of TB. Out of the 140 sputum specimens, 38 (27.14%) were positive by ZN/AFB smear microscopy, 60 (42.86%) by the GeneXpert MTB/RIF assay, and 41 (29.28%) by MTB culture on LJ medium. With respect to the detection of bacillary load, out of the sixty specimens that tested positive for bacillary load, the semi-quantitative results showed that 11 samples (18.3%) had high results (mean cycle threshold (CT) value =14.3), 25 (41.7%) had medium results (mean CT value =20.8), 15 (25.0%) had low results (mean CT value = 25.4), and 9 (15.0%) had very low results (mean CT value =30.1).Rifampicin resistance was detected in 7 out of 60 (11.67%) of the MTB-positive specimens, while 53 (88.33%) showed no rifampicin resistance (Rif-S).
Conclusion: The GeneXpert MTB/RIF assay demonstrates high sensitivity and specificity, making it a valuable diagnostic tool for the rapid detection of TB bacilli and the simultaneous identification of RR-TB. Early detection of rifampicin resistance is crucial in curbing the spread of multidrug-resistant TB (MDR-TB) strains.
References
Alame-Emane, A. K., Pierre-Audigier, C., Aboumegone-Biyogo, O. C., Nzoghe-Mveang, A., Cadet-Daniel, V., Sola, C., ... and Takiff, H. E. (2017). Use of GeneXpert remnants for drug resistance profiling and molecular epidemiology of tuberculosis in Libreville, Gabon. Journal of Clinical Microbiology, 55(7): 2105-2115.
Aljanaby, A. A. J., Al-Faham, Q. M. H., Aljanaby, I. A. J., and Hasan, T. H. (2022). Epidemiological study of mycobacterium tuberculosis in Baghdad governorate, Iraq. Gene Reports, 26: 101467.
Al-Mussawi, A. A., Ali, N. H., and Abed, A. H. (2017). Molecular quantification of Rifampicin-resistance in Mycobacterium tuberculosis. Egyptian Journal of Chest Diseases and Tuberculosis, 66(3): 425-427.
AL-Obaidy, M. W. (2013). A Prevelance of Drug-Resistance in Previously Treated Tuberculous Patients in Baghdad. Iraqi Postgraduate Medical Journal, 12 :626-631.
Atehortúa, S., Ramírez, F., Echeverri, L. M., Peñata, A., and Ospina, S. (2015). Xpert MTB/RIF test performance assay in respiratory samples at real work settings in a developing country. Biomédica, 35(1): 125-130.
Blakemore, R., Nabeta, P., Davidow, A. L., Vadwai, V., Tahirli, R., Munsamy, V., ... and Alland, D. (2011). A multisite assessment of the quantitative capabilities of the Xpert MTB/RIF assay. American Journal of Respiratory and Critical Care Medicine, 184(9), 1076-1084.
Boakye-Appiah, J. K., Steinmetz, A. R., Pupulampu, P., Ofori-Yirenkyi, S., Tetteh, I., Frimpong, M., ... and Phillips, R. O. (2016). High prevalence of multidrug-resistant tuberculosis among patients with rifampicin resistance using GeneXpert Mycobacterium tuberculosis/rifampicin in Ghana. International Journal of Mycobacteriology, 5(2), 226-230.
Bodmer, T., and Ströhle, A. (2012). Diagnosing pulmonary tuberculosis with the Xpert MTB/RIF test. Journal of Visualized Experiments, (62), e3547.
Boehme, C. C., Nabeta, P., Hillemann, D., Nicol, M. P., Shenai, S., Krapp, F., ... & Perkins, M. D. (2010). Rapid molecular detection of tuberculosis and rifampin resistance. New England Journal of Medicine, 363(11), 1005-1015.
Caulfield, A. J., and Wengenack, N. L. (2016). Diagnosis of active tuberculosis disease: From microscopy to molecular techniques. Journal of Clinical Tuberculosis and Other Mycobacterial Diseases, 4: 33-43.
Dagnra, A. Y., Mlaga, K. D., Adjoh, K., Kadanga, E., Disse, K., and Adekambi, T. (2015). Prevalence of multidrug-resistant tuberculosis cases among HIV-positive and HIV-negative patients eligible for retreatment regimen in Togo using GeneXpert MTB/RIF. New Microbes and New Infections, 8: 24-27.
David, S., Katalinić-Janković, V., and Cirillo, D. (2018). 4. Smear microscopy. In Handbook on tuberculosis laboratory diagnostic methods in the European Union– Updated 2018. Stockholm: ECDC.
Desissa, F., Workineh, T., and Beyene, T. (2018). Risk factors for the occurrence of multidrug-resistant tuberculosis among patients undergoing multidrug-resistant tuberculosis treatment in East Shoa, Ethiopia. BMC Public Health, 18(1): 1-6.
Dorman, S. E., Chihota, V. N., Lewis, J. J., Shah, M., Clark, D., Grant, A. D., ... & Churchyard, G. J. (2012). Performance characteristics of the GeneXpert MTB/RIF assay in a tuberculosis prevalence survey. PloS one, 7(8), e43307.
Dzodanu, E. G., Afrifa, J., Acheampong, D. O., and Dadzie, I. (2019). Diagnostic yield of fluorescence and Ziehl-Neelsen staining techniques in the diagnosis of pulmonary tuberculosis: A comparative study in a district health facility. Tuberculosis Research and Treatment, 2019: 4091937.
El-Hajj, H. H., Marras, S. A., Tyagi, S., Kramer, F. R., and Alland, D. (2001). Detection of rifampin resistance in Mycobacterium tuberculosis in a single tube with molecular beacons. Journal of Clinical Microbiology, 39(11), 4131-4137.
Fouda, M. E., Eman, R., Gawad, A., Sahar, M., and Fayed, M. H. (2019). A study of the added value of Xpert MTB/RIF assay for assessment of pulmonary tuberculosis transmission risk. Egyptian Journal of Medical Microbiology,28(3): 141-8.
Fradejas, I., Ontañón, B., Muñoz-Gallego, I., Ramírez-Vela, M. J., and López-Roa, P. (2018). The value of xpert MTB/RIF-generated CT values for predicting the smear status of patients with pulmonary tuberculosis. Journal of Clinical Tuberculosis and Other Mycobacterial Diseases, 13: 9-12.
Geleta, D. A., Megerssa, Y. C., Gudeta, A. N., Akalu, G. T., Debele, M. T., and Tulu, K. D. (2015). Xpert MTB/RIF assay for diagnosis of pulmonary tuberculosis in sputum specimens in a remote health care facility. BMC Microbiology, 15(1): 1-6.
Gilpin, C., Korobitsyn, A., Migliori, G. B., Raviglione, M. C., and Weyer, K. (2018). The World Health Organization standards for tuberculosis care and management. European Respiratory Journal, 51(3).
Habte, D., Melese, M., Hiruy, N., Gashu, Z., Jerene, D., Moges, F., ... and Tessema, B. (2016). The additional yield of GeneXpert MTB/RIF test in the diagnosis of pulmonary tuberculosis among household contacts of smear positive TB cases. International Journal of Infectious Diseases, 49: 179-184.
Helb, D., Jones, M., Story, E., Boehme, C., Wallace, E., Ho, K., ... & Alland, D. (2010). Rapid detection of Mycobacterium tuberculosis and rifampin resistance by use of on-demand, near-patient technology. Journal of Clinical Microbiology, 48(1), 229-237.
Horne, D. J., Narita, M., Spitters, C., Parimi, S., & Dodson, D. (2010). Quality of care in tuberculosis treatment: improving adherence to guidelines. Journal of Clinical Tuberculosis and Other Mycobacterial Diseases, 1(1), 4-10.
Kaewseekhao, B., Nuntawong, N., Eiamchai, P., Roytrakul, S., Reechaipichitkul, W., and Faksri, K. (2020). Diagnosis of active tuberculosis and latent tuberculosis infection based on Raman spectroscopy and surface-enhanced Raman spectroscopy. Tuberculosis, 121: 101916.
Lawn, S. D., Mwaba, P., Bates, M., Piatek, A., Alexander, H., Marais, S., ... & McNerney, R. (2011). Advances in tuberculosis diagnostics: the Xpert MTB/RIF assay and future prospects for a point-of-care test. The Lancet Infectious Diseases, 11(5), 349-361.
Masenga, S. K., Mubila, H., and Hamooya, B. M. (2017). Rifampicin resistance in mycobacterium tuberculosis patients using GeneXpert at Livingstone Central Hospital for the year 2015: a cross sectional explorative study. BMC Infectious Diseases, 17(1): 1-4.
Mekonnen, F., Tessema, B., Moges, F., Gelaw, A., Eshetie, S., and Kumera, G. (2015). Multidrug resistant tuberculosis: prevalence and risk factors in districts of metema and west armachiho, Northwest Ethiopia. BMC Infectious Diseases, 15(1): 1-6.
Mulisa, G., Workneh, T., Hordofa, N., Suaudi, M., Abebe, G., and Jarso, G. (2015). Multidrug-resistant Mycobacterium tuberculosis and associated risk factors in Oromia Region of Ethiopia. International Journal of Infectious Diseases, 39: 57-61.
Munir, M. K., Rehman, S., Aasim, M., Iqbal, R., and Saeed, S. (2015). Comparison of Ziehl Neelsen microscopy with GeneXpert for detection of Mycobacterium tuberculosis. IOSR Journal of Dental and Medical Sciences, 14(11): 56-60.
Najjingo, I., Muttamba, W., Kirenga, B. J., Nalunjogi, J., Bakesiima, R., Olweny, F., ... and Ssengooba, W. (2019). Comparison of GeneXpert cycle threshold values with smear microscopy and culture as a measure of mycobacterial burden in five regional referral hospitals of Uganda-A cross-sectional study. PLoS One, 14(5): e0216901.
Rasheed, W., Rao, N. A., Adel, H., Baig, M. S., and Adil, S. O. (2019). Diagnostic accuracy of Xpert MTB/RIF in sputum smear-negative pulmonary tuberculosis. Cureus, 11(8).
Riaz, M., Mahmood, Z., Javed, M. T., Javed, I., Shahid, M., Abbas, M., and Ehtisham-ul-Haque, S. (2016). Drug resistant strains of Mycobacterium tuberculosis identified through PCR-RFLP from patients of Central Punjab, Pakistan. International Journal of Immunopathology and Pharmacology, 29(3): 443-449.
Sambarey, A., Devaprasad, A., Mohan, A., Ahmed, A., Nayak, S., Swaminathan, S., ... and Chandra, N. (2017). Unbiased identification of blood-based biomarkers for pulmonary tuberculosis by modeling and mining molecular interaction networks. EBioMedicine, 15: 112-126.
Shi, J., Dong, W., Ma, Y., Liang, Q., Shang, Y., Wang, F., ... and Pang, Y. (2018). GeneXpert MTB/RIF outperforms mycobacterial culture in detecting Mycobacterium tuberculosis from salivary sputum. BioMed Research International, 2018: 1514381.
Singh, U. B., Pandey, P., Mehta, G., Bhatnagar, A. K., Mohan, A., Goyal, V., ... and Samantaray, J. C. (2016). Genotypic, phenotypic and clinical validation of GeneXpert in extra-pulmonary and pulmonary tuberculosis in India. PloS one, 11(2), e0149258
Steingart, K. R., Schiller, I., Horne, D. J., Pai, M., Boehme, C. C., & Dendukuri, N. (2014). Xpert® MTB/RIF assay for pulmonary tuberculosis and rifampicin resistance in adults. Cochrane Database of Systematic Reviews, (1).
Wang, W. H., Takeuchi, R., Jain, S. H., Jiang, Y. H., Watanuki, S., Ohtaki, Y., ... and Ito, E. (2020). A novel, rapid (within hours) culture-free diagnostic method for detecting live Mycobacterium tuberculosis with high sensitivity. EBioMedicine, 60: 103007.
World Health Organization (WHO). (2013). Automated real-time nucleic acid amplification technology for rapid and simultaneous detection of tuberculosis and rifampicin resistance: Xpert MTB/RIF assay for the diagnosis of pulmonary and extrapulmonary TB in adults and children. Policy update. WHO Guidelines .
World Health Organization. (2014). Mycobacteriology laboratory manual. Global Laboratory Initiative Advancing TB Diagnosis. Geneva, Switzerland: WHO.
Zaw, M. T., Emran, N. A., and Lin, Z. (2018). Mutations inside rifampicin-resistance determining region of rpoB gene associated with rifampicin-resistance in Mycobacterium tuberculosis. Journal of Infection and Public Health, 11(5): 605-610.