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Aims: To differentiate and Characterize Mycobacterium tuberculosis complex causing pulmonary tuberculosis across North Central Nigeria
Study Design: This was a simple descriptive health-based study that involved clinically suspected tuberculosis patients who were referred to two selected General Hospitals for diagnosis in each of the states across North Central Nigeria.
Place and Duration of Study: This study was carried out in the North Central zone of Nigeria and it included all the seven States across North Central Nigeria using two General Hospitals per state. The study included 371 positive sputum samples drawn from 2800 suspected pulmonary TB patients between 2017 and 2018.
Methodology: A total of 371 GeneXpert positive sputa for TB were decontaminated using N Acetyl Cysteine and Sodium Hydroxide under a level 3 Bio-safety cabinet and the resulting sediment was cultured on Lowestein- Jensen (LJ) media containing glycerol and pyruvate at 37ºC in a slanted position, SD- Bioline (TB Ag MPT 64) for the differentiation of MTBC from NTM was carried out using the isolates from LJ culture. Evaluation of speciation was done using Line Probe Assay to determine the predominant species of MTBC. All the protocols used in this study followed the manufacturer’s manual strictly.
Results: A total of 371 decontaminated positive GeneXpert sputa derived from 2800 suspected PTB participants were cultured on Lowestein- Jensen (LJ) medium and 302(81.40%) was found positive while 69(18.60%) were found negative. Out of the culture positive isolates, 288 (95.36%) isolates were detected on SD-BIOLINE TB Ag MPT 64 ® assay for MTBC and 14 (4.64%) as NTM. Of the 288 MTBC, three different species were identified; 272 (94.64%) were M. tuberculosis/M. Canetti, 7 (2.43%) were M. africanum and 9(3.13) showed a no MTBC reaction band on all the samples that were analysed.
Conclusion: Differentiations of MTBC from NTM has help to re-confirm that not all symptoms of pulmonary infection are caused by MTBC but NTM are implicated due to their distribution in the environment, however, molecular characterisation method has narrow our findings down to M. tuberculosis/M. canettii as the predominant specie of MTBC circulating in the region, although, M. africaum was also detected and these two species of MTBC are the leading cause of pulmonary tuberculosis across all the North Central state of Nigeria.
World Health Organization. Global tuberculosis report; 2015.
[Accessed on 9 September 2016]
Centre Disease Control. Reported Tuberculosis in the United States, Atlanta. Ga: U.S. Department of Health and Human Services, CDC; 2012.
Grace Wambura Mbuthia, Charles Owour, OlungahTom, Gesora Ondicho. Knowledge and perceptions of tuberculosis among patients in a pastoralist community in Kenya: A qualitative study. Pan African Medical Journal. 2018;287:1-6.
World Health Organization. Global tuberculosis report; 2018.
(Accessed September 18, 2018)
World Health Organization. Global tuberculosis report. (WHO/HTM/ TB/2017.23). Geneva: WHO; 2017.
(Accessed 21 June 2018)
Ebonyi AO, Oguche S, Agbaji OO, Sagay AS, Okonkwo PI, Idoko JA, Kanki PJ. Mortality among pulmonary tuberculosis and HIV-1 co-infected Nigerian children being treated for pulmonary tuberculosis and on antiretroviral therapy: A retrospective cohort study. GERMS. 2016; 6(4):139-150.
Lienhardt C, Glaziou, P Uplekar. Global tuberculosis control: lessons learnt and future prospects. Nat Rev Microbiol. 2012; 10:407-16.
Sandgren A, Schepisi, MS Sotgiu. Tuberculosis transmission between foreign- and native-born populations in the EU/EEA: A systematic review. Eur Respir J. 2014;43:1159-71.
Peters JS, Calder B, Gonnelli G, Degroeve S, Rajaonarifara E, Mulder N. Identification of quantitative proteomic differences between Mycobacterium tuberculosis lineages with altered virulence. Front Microbiol. 2016;7:813.
Moreno V, Espinoza B, Barley K, Paredes M, Bichara D, Mubayi A. The role of mobility and health disparities on the transmission dynamics of tuberculosis. Theor Biol Med Model. 2017;14:3.
Schürch AC, Van Soolingen D. DNA fingerprinting of Mycobacterium tuber-culosis: From phage typing to whole-genome sequencing. Infection Genetics and Evolution. 2012;12(4):602–609.
Wikipedia. The free encyclopedia; 2019.
(Accessed, 22 of April 2019)
Yamane Taro. Statistics, An Introductory Analysis. 3rd Ed., New York: Harper and Row; 1973.
World Health Organization. Guideline for treatment of drug-susceptible tuberculosis and patients care: Updated edition. (WHO/HTM/ TB/2017.23). Geneva: WHO; 2017b.
(Accessed 21 June2018)
Joshua OO, Gospel TO, Emeka UE, Chukwukere E. Nigerian National TB standard operating procedures manual for Laboratories. National Tuberculosis and Leprosy Control program (NTBLCP). Ministry of Health, Nigeria and American Society for Microbiology, 1st edition. 2013; 1-244.
Centre for Disease Control. Guidelines for safe work practices in human and animal medical diagnostic laboratories: Recommendations for CDC-Convened Biosafety Blue-Ribbon Panel. Southern China. Molecular Biology Reports. 2012; 38(3):2185–2192.
Yates M, Drobniewski F, Wilson S. Evaluation of a rapid PCR-based epidemiological typing method for routine studies of Mycobacterium tuberculosis. J Clin Microbiol. 2002; 40:712–4.
Ibrahim S, Abubakar UB, Danbirni S, Usman A, Ballah FM, Kudi CA, Lawson L, Abdulrazak GH, Abdulkadir IA. The Use of SD Bioline TB AgMPT64; Detection Assay for Rapid Characterization of Mycobacteria in Nigeria. 2017;11(2):94-100
Richter E, Weizenegger M, Fahr AM, Rüsch-Gerdes S. Usefulness of the GenoType MTBC assay for differentiating species of the Mycobacterium tuberculosis complex in cultures obtained from clinical specimens. J. Clin. Microbiol. 2004;42: 4303–4306.
Somoskovi A, Dormandy J, Rivenburg J. Direct comparison of the GenoType MTBC and genomic deletion assays in terms of ability to distinguish between members of the Mycobacterium tuberculosis complex in clinical isolates and in clinical specimens. J. Clin. Microbiol. 2008;46:1854–1857.
Wagner D, Young L. Non-tuberculous mycobacterial infections: A clinical review. Infection. 2004;32:257–70.
Shaarawy H, Elhawary AT. Risk factors for atypical mycobacterial disease in patients with smear positive pulmonary TB. Egypt J Chest Dis Tuberc. 2014;63:657-61.
Shahraki AH, Heidarieh P, Bostanabad SZ, Khosravi AD, Hashemzadeh M, Khandan S. Multidrug-resistant tuberculosis may be nontuberculous mycobacteria. Eur J Intern Med. 2015;26:279-84.
Aliyu G, El-Kamary SS, Abimiku A, Brown C, Tracy K, Hungerford L. Prevalence of non-tuberculous mycobacterial infections among tuberculosis suspects in Nigeria. Plos One. 2013;8(5):e63170.
van Halsema CL, Chihota VN, Gey van Pittius NC, Fielding KL, Lewis JJ, van Helden PD. Clinical relevance of nontuberculous mycobacteria isolated from sputum in a gold mining workforce in South Africa: An observational, clinical study. Biomed Res Int. 2015;959:107.
Mertaniasih NM, Kusumaningrum D, Koendhori EB, Soedarsono, Kusmiati T, Dewi DN. Nontuberculous mycobacterial species and Mycobacterium tuberculosis complex coinfection in patients with pulmonary tuberculosis in Dr. Soetomo Hospital, Surabaya, Indonesia. Int J Mycobacteriol. 2017;6:9-13.
Shao Y, Chen C, Song H, Li G, Liu Q, Li Y. The epidemiology and geographic distribution of nontuberculous mycobacteria clinical isolates from sputum samples in the eastern region of China. PLoS Negl Trop Dis. 2015;9:e0003623.
Singh AK, Maurya AK, Umrao J, Kant S, Kushwaha RAS, Nag VL, Dhole, TN. Role of genotype® mycobacterium common mycobacteria/additional species assay for rapid differentiation between Mycobacterium tuberculosis complex and different species of non-tuberculous mycobacteria. Journal of laboratory Physicians. 2013;5(2):83.
Gagneux S, DeRiemer K, Van T, Kato-Maeda M, de Jong BC. Variable host-pathogen compatibility in Mycobacterium tuberculosis. Proc Natl Acad Sci USA. 2006;103:2869-2873.
Wirth T, Hildebrand F, Allix-Béguec C, Wölbeling F, Kubica T, Kremer K, van Soolingen D, Rüsch-Gerdes S, Locht C, Brisse S, Meyer A, Supply P, Niemann S. "Origin, spread and demography of the Mycobacterium tuberculosis complex. PLoS Pathog. 2008;4(9):e1000160.
DOI: 10.1371/journal.ppat. 1000160. PMC 2528947.
Geoffrey Arasa Ouno, Rose Kakai, Henry ND, Nyamogoba Biegon RK, Cornelius K. Magut, Wilfred B. Murithi. Molecular identity of mycobacteria isolates in new cases of pulmonary tuberculosis patients in Kisumu County, Western Kenya. Journal of Natural Sciences Research. 2017; 20:2225-0921.
Niobe-Eyangoh SN, Kuaban C, Sorlin P. Genetic biodiversity of Mycobacterium tuberculosis complex species from patients with pulmonary tuberculosis in Cameroon. Journal of Clinical Microbiology. 2013; 41(6):2547–2553.
Mbugi EV, Katale BZ, Siame KK, Keyyu J D, Kendall SL, Dockrell HM, Warren RM. Genetic diversity of Mycobacterium tuberculosis isolated from tuberculosis patients in the Serengeti ecosystem in Tanzania. Tuberculosis. 2015;5(2):170-178.
Ndadilnasiya Endie Waziri, Simeon Cadmus, Patrick Nguku, Olufunmilayo Fawole, Olajide Adewale Owolodun, Hyelshilni Waziri, Luka Ibrahim, Oladayo Biya, Saheed Gidado, Samuel Badung, Peterside Kumbish, Peter Nsubuga. Factors associated with tuberculosis among patients attending a treatment center in Zaria, North-west Nigeria, 2010. Pan Afr Med J. 2014;18(Supp 1):5.
Sani RA, Garba SA, Oyeleke SB, Abalaka ME. Prevalence of pulmonary tuberculosis (PTB) in Minna and Suleja Niger State, Nigeria. American Journal of Medicine and Medical Sciences. 2015;5(6):287-291.
Figueiredo EES, Silvestre FG, Campos WN, Furlanetto LV. Identification of Mycobacterium bovis isolates by a multiplex PCR. Brazilian Journal of Microbiology. 2010;41(2):231-233.