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1. Mycobacterium tuberculosis Complex
Genotype Diversity and Drug Resistance
Profiles in a Pediatrics Population in,
Addis Ababa Ethiopia
Sebsib Neway
October, 2022
1

2. Out line
• Background
• Objective
• Statement Problem
• Objective
• Methodology
• Reference
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3. Background
• Tuberculosis (TB) remains a major global public health
problem and is the leading cause of death from a single
bacterium, MTB (Leopold D et al., 2015).
• It accounts for more than 10% of hospital admissions and
death in children in most developing African’s countries
(Bereket et al., 2013).
• The high burden of TB among children is assumed to be due to
the high prevalence of TB in adults, HIV co-infection,
malnutrition and other conditions related to poverty (Richard Copin
et al., 2016). 3

4. Cont.
• It remains one of the most threatening infectious diseases,
causing 10.4 million incident cases and 1.4 million deaths
per year.
• Children rarely transmit the disease, and thus contribute
little to the maintenance of the TB epidemic (Roberto
Zenteno-Cuevas et al., 2021).
• According to the WHO, at least half a million children
become ill with TB (Robindra et al., 2019)
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5. Cont.
• Traditionally, pediatric tuberculosis has been relatively
neglected, although recent years have seen a welcome increase
in policy focus including the goal of zero childhood
tuberculosis deaths (Bereket et al., 2013).
• Strain related influence on pulmonary TB may potentially also
impact on TB transmission (Tekle et al., 2020).
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6. Cont.
• There are limited data regarding strain associated differences
in the host-pathogen interaction in children, as the
paucibacillary nature of TB in young children makes
bacteriological confirmation and mycobacterial genotyping
challenging (Mercedes et al., 2011).
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7. Cont.
• The emergence of MDR tuberculosis and extensively XDR
tuberculosis defined as resistant to Isoniazid and Rifampicin,
• In addition to any fluoroquinolones, and at least one injectable
anti-tuberculosis drug), has worsened tuberculosis morbidity
and mortality.
• There is no adequate data is available in Ethiopia and even
limited worldwide studies regarding to the children population.
• Therefore, the present study was conducted to determine
genotype diversity and drug resistance profiles anti-TB drug
resistance in a pediatric population in Addis Ababa. 7

8. Objective
• To determine the genetic diversity of M. tuberculosis strains in
pediatrics patients in Addis Ababa, Ethiopia.
• To assess the prevalence of MTB and drug resistant pattern in
children
• To determine the frequency of drug resistance in both
genotype and phenotype DST and comparative results
• To determine the associate factors
• To assess treatment outcomes
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9. Methodology
Study design
• A cross-sectional study will be applied.
• M tuberculosis isolates collected from Gastric lavage or sputum
samples TB patients will be further processed and inoculated.
• It will be proceed from growth TB bacteria for phenotypic and
genotypic drug resistance testing using BACTEC 960, MIGIT,
Spologotyping and molecular drug resistance using WGS.
• A total of 366 study participants will be recruited.
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10. Statement of Problem
• TB remains the most devastating infectious disease,
particularly in Ethiopia.
• Still limited data available on Pediatrics populations
• Identification of MTBC requires confirmation by molecular
tools with a better discriminatory i.e. sequencing
• Highly specific and sensitive diagnostic method will used
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11. Sampling
• Sampling Consecutive newly diagnosed pulmonary TB Pediatrics patients from
selected health facilities (Hospitals) found in Addis Ababa will be recruited
until 366 samples recovered.
• At least 5ml Gastric lavage/sputum specimens will be collected.
• Specimens will be further processed with 3% NALC-NaOH concentration
method and inoculated on LJ culture and Bactec 960 MGIT culture media.
• Culture grown Isolates will be further analyzed for drug resistance tasting on
MGIT first and line drugs kit).
• DNA will be extracted from fresh isolates that will be resistant for any of the
drugs using fast DNA kit (MP Biomedicals) for WGS of isolates at AHRI
laboratory. 11

12. Cont.
• MTB isolates resistant for first line drugs will be further subjected to
second line DST.
• Data Analysis Data will be checked, cleared and entered into AHRI
data management center. Proportions, odds ratio, binary and
multiple logistic regressions will be used to assess the presence and
degree of association between the study variables.
• The WGS data will be further investigated and verified for mutation
sites associated with resistance using the appropriate bioinformatics
tools.
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13. Cont.
• Ethical considerations: will be approved by the Institutional
Research Ethics Board of Addis Ababa University, College of
Natural and Computational Science and AHRI/ALERT
institutional ethical review committee.
• Informed written and signed consent will be obtained from
each participant before data collection.
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14. Cont.
• Confidentiality of participants’ data will be maintained by
coding the samples and data collection sheets.
• Sample Size: Estimated using formula N =Z α /2 P(1-
P)/d2=366 with addition to 10% The prevalence was taken
from study done in Jimma 2013 i.e. 31.7%
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15. Eligibility Criteria
• Inclusion:
-All pediatric patient newly diagnosed for MTB
• Exclusion: Those
-Pediatric patient under treatment and recent time
contact with MTB drugs
-Pediatric families refused for consent
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16. Budget
• Budget will be from different AHRI Project sources or another
funder organization
• The manuscript will be published cooperating with University on
international journal
• The Abstract will be presented in different scientific conference
Opportunity
• The project will be done AHRI
• Senior and lead researchers are found in AHRI
• Sample sources sites very well since already linked with AHRI
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17. Reference.
1. Genetic diversity and drug resistance pattern of Mycobacterium tuberculosis strains isolated from pulmonary
tuberculosis patients in the Benishangul Gumuz region and its surroundings, Northwest Ethiopia
2. Genotype diversity of Mycobacterium isolates from children in Jimma, Ethiopia
3. Micro bead-based spoligotyping of Mycobacterium tuberculosis from Ziehl-Neelsen-stained microscopy preparations
in Ethiopia
4. Mycobacterium tuberculosis Complex Genotype Diversity and Drug Resistance Profiles in a Pediatric Population in
Mexico
5. Phylogenetic associations with drug-resistant Mycobacterium tuberculosis isolates in a pediatrics population
6. Hesseling AC, Marais BJ, Kirchner HL, Mandalakas AM, Brittle W, Victor TC, Warren RM, Schaaf HS.
Mycobacterial genotype is associated with disease phenotype in children. The International journal of tuberculosis and
lung disease. 2010 Oct 1;14(10):1252-8. 7.
7. Hove P, Molepo J, Dube S, Nchabeleng M. Genotypic diversity of Mycobacterium tuberculosis in Pretoria. Southern
African Journal of Epidemiology and Infection. 2012 Jan 1;27(2):77-83
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18. Thank you
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19. References
• Genetic diversity and drug resistance pattern of Mycobacterium
tuberculosis strains isolated from pulmonary tuberculosis patients in the
Benishangul Gumuz region and its surroundings, Northwest Ethiopia
• Genotype diversity of Mycobacterium isolates from children in Jimma,
Ethiopia
• Micro bead-based spoligotyping of Mycobacterium tuberculosis from
Ziehl-Neelsen-stained microscopy preparations in Ethiopia
• Mycobacterium tuberculosis Complex Genotype Diversity and Drug
Resistance Profiles in a Pediatric Population in Mexico
• Phylogenetic associations with drug-resistant Mycobacterium tuberculosis
isolates in a pediatrics population
• Hesseling AC, Marais BJ, Kirchner HL, Mandalakas AM, Brittle W, Victor
TC, Warren RM, Schaaf HS. Mycobacterial genotype is associated with
disease phenotype in children. The International journal of tuberculosis and
lung disease. 2010 Oct 1;14(10):1252-8.
• 7. Hove P, Molepo J, Dube S, Nchabeleng M. Genotypic diversity of
Mycobacterium tuberculosis in Pretoria. Southern African Journal of
Epidemiology and Infection. 2012 Jan 1;27(2):77-83.
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