WORLD TB DAY COMMEMORATED ON 24TH MARCH OF.pptx

MTB Complex Resistance Profile and Genotyping

1. WORLD TB DAY COMMEMORATED ON 24 TH MARCH OF EVERY YEAR By Amb . Dr. Hamisu Umar Takalmawa , PhD, FMLSCN, MASM, MNSM.

2. Each year, stakeholders recognize World TB Day on March 24 . NHC , Dar es Salaam is joining the World to commemorate this day today . The event commemorates the date in 1882 when Dr . Robert Koch, Father of Bacteriology announced his discovery of Mycobacterium tuberculosis This discovery opened the way towards diagnosing and curing this disease . World TB Day 2023 focuses on the theme, ‘Yes! We can end TB !’ T o raise public awareness about the devastating health , social and economic consequences of TB, and to scale up efforts to end the global epidemic.

3. Builds on previous efforts and commitments by various National Governments to invest resources to ramp up the fight against TB The commemoration comes amidst renewed efforts to fight the spread of the COVID-19 that has put ending global TB at risk T o ensure equitable access to prevention and care in line with WHO’s drive towards achieving Universal Health Coverage, (UHC) H ealth promotion, Prevention, Treatment, Rehabilitation and Palliative care Every country has its own path to achieving UHC and decides what to cover based on the needs of their people and the resources at hand. However access to health services and information are universal basic human right .

4. The spotlight of World TB Day 2023 is on urging countries to ramp up progress in the lead-up to the UN High-Level Meeting on TB. It is expected that WHO will also issue a call to action with partners urging Member States to accelerate the rollout of the new WHO-recommended shorter all-oral treatment regimens for drug-resistant tuberculosis (DR-TB).

5. TUBERCULOSIS I s a potentially serious ID that mainly affects the respiratory tract (PTB) but other organs of the body could be affected (Extra-Pulmonary TB) S preads from person to person through tiny droplets released into the air when an infected and diseased person coughs and/or sneezes, talk or even laugh Figure 1.0: How Tuberculosis is transmitted

6. GLOBAL TB TREATMENT COVERAGE With currently-recommended treatments (4–6 months course of anti-TB drugs), about 85% of people can be cured. Regimens of 1–6 months are available to treat TB infection Globally in 2021, TB treatment coverage (for both HIV-negative and HIV-positive people) was 61% Up from 58% in 2020 but substantially lower than 69% in 2019

7. TB remains the world’s most deadly ID that claims > 1x 10 6 lives each year and affects millions more, with enormous social, economic and psychological impacts on families and communities . Until the Coronavirus (COVID-19) pandemic, TB was the leading cause of death from a single infectious agent , ranking above HIV/AIDS . TB is the 13th leading cause of death worldwide . It was also the leading killer of people with HIV and a major cause of deaths related to antimicrobial resistance .

8. GLOBAL FIGURE Globally , the number of people newly diagnosed and officially reported with TB fell from 7.1 million in 2019 to 5.8 million in 2020 (-18%), with a partial recovery to 6.4 million in 2021 Nigeria (4.4 %) and Democratic Republic of the Congo (2.9%) India (28%), Indonesia (9.2%), China (7.4%), the Philippines (7.0%), Pakistan (5.8%), Bangladesh (3.6 %)

9. Figure 2: Countries with High TB in 2021 that accounted for > 2/3 of global cases

10. Three high TB burden countries have reached or passed the first milestones of the End TB Strategy for both reductions in TB incidence and TB deaths: Kenya (in 2018), Tanzania (in 2019) and Zambia (in 2021 ). Ethiopia is very close .

11. TB IN NIGERIA Despite TB being a vaccine-preventable disease, statistics from the WHO shows that every year, about 245,000 Nigerians die from the disease, and about 590,000 new cases occur. The data further shows that around 140,000 of this figure are also HIV-positive . Incidence of tuberculosis (per 100,000 people) in Nigeria was reported at 219 in 2021 , according to the World Bank collection of development indicators , compiled from officially recognized sources.

12. Figure 3 .0 TB cases in Nigeria between 2010 and 2020

13. Figure 4: TB –HIV Co-infection in Nigeria

14. MOLECULAR EPIDEMIOLOGY OF MTB COMPLEX IN KANO, NIGERIA SIGNIFICANCE OF THE STUDY There was no data on MTB genotypes, their diversity/distribution and resistance pattern in Kano No study worked to identify or recognize any MTB lineage/family/variant circulating in Kano i.e. no base line genotyping information despite increase in the incidence of DR-TB ( MDR and potential XDR) ( Maiyaki et al, 2017; Mohammed et al, 2017 ). To have a directed approach in treatment modalities

15. Molecular epidemiology (ME) that employs DNA typing methods is one of the main areas in TB research which is widely used to study the transmission dynamics of TB Based on the various polymorphism in the genome that is used as genetic marker. The three genotyping tools widely used to differentiate various strains and study the evolutionary relationships between them are IS6110-RFLP, Spoligotyping , and MIRU-VNTR (Braden et al . 1997, Kammerbeek et al , 1998 and Benjamin et al 2012).

16. Figure 5 : Hypothetical genome of MTB strain X and polymorphic repetitive sequences such as IS6110, DR and MIRUs for genotyping. Adopted from Supply et al., 2006

17. Genetic Diversity, Resistance Pattern and Transmission Dynamics of Mycobacterium tuberculosis Complex in Kano, North-West Nigeria. Hypotheses The MTB complex strains circulating in Kano are not genotypically diverse (same genotypes ) ii. MTB complex genotypes circulating in Kano are not associated with any drug resistance

18. The knowledge about local strains is crucial: Successful treatment of the disease relies on good characterisation of these strains and a more directed approach in prevention and control policies – patients, clinicians (IDS/Pulmonologists ) and public health stakeholders.

19. Figure 7: GeneXpert MTB/RIF assay workflow for rapid diagnosis test of MTB and Rifampicin resistance. 

20. Temperature Time Clinical specimens Cultured specimen 95°C 15min 1 cycle 1 cycle 95°C 65°C 30 seconds 2 minutes 20 cycles 10 cycles 95°C 50°C 70°C 25 seconds 40 seconds 40 seconds 30 cycles 20 cycles 70°C 8 minutes 1 cycle 1 cycle Amplicon storage at +8 to – 20°C Table 1 : Amplification profile for first and second line anti-TB drug LPA

21. Alias VNTR loci Primer sequence Mtub04 424 CTTGGCCGGCATCAAGCGCATTATT GGCAGCAGAGCCCGGGATTCTTC MIRU4 (ETR-D) 580 GCGCGAGAGCCCGAACTGC GCGCAGCAGAAACGCCAGC MIRU10 960 GTTCTTGACCAACTGCAGTCGTCC GCCACCTTGGTGATCAGCTACCT MIRU16 1644 TCGGTGATCGGGTCCAGTCCAAGTA CCCGTCGTGCAGCCCTGGTAC Mtub21 1955 AGATCCCAGTTGTCGTCGTC CAACATCGCCTGGTTCTGTA QUB26 4052 AACGCTCAGCTGTCGGAT CGGCCGTGCCGGCCAGGTCCTTCCCGAT Mtub30 2401 CTTGAAGCCCCGGTCTCATCTGT ACTTGAACCCCCACGCCCATTAGTA MIRU26 2996 TAGGTCTACCGTCGAAATCTGTGAC CATAGGCGACCAGGCGAATAG Mtub39 3690 CGGTGGAGGCGATGAACGTCTTC TAGAGCGGCACGGGGGAAAGCTTAG MIRU31 (ETR-E) 3192 ACTGATTGGCTTCATACGGCTTTA GTGCCGACGTGGTCTTGAT QUB4156 4156 TGACCACGGATTGCTCTAGT GCCGGCGTCCATGTT MIRU40 802 GGGTTGCTGGATGACAACGTGT GGGTGATCTCGGCGAAATCAGATA ETR-A 2165 AAATCGGTCCCATCACCTTCTTAT CGAAGCCTGGGGTGCCCGCGATTT QUB11b 2163b CGTAAGGGGGATGCGGGAAATAGG CGAAGTGAATGGTGGCAT ETR-C 577 CGAGAGTGGCAGTGGCGGTTATCT AATGACTTGAACGCGCAAATTGTGA Table 2: Original 15 loci consisted of 5 exact tandem repeats (ETR A-E) and 10 MIRUs

22. Figure 8: Experimental workflow of MIRU-VNTR analysis of MTBC strains in Kano, Nigeria

23. RESULTS AND DISCUSSIONS

24. Figure : Dynamics of TB among patients attending IDH , Kano from 2015-2018

25. Figure 12: Type of TB and treatment outcome in Kano

26. Since there is consistent rise of prevalence from 2015 to 2018, the study can conclude that DOTS clinic is improving the detection rate of TB and consequently playing a big supervisory role in TB treatment and control. The IDH TB treatment unit is successfully implementing the WHO’s DOTS strategy and has achieved good treatment completion rates in recent years

27. Figure : Prevalence of Extra-pulmonary TB among presumptive cases in IDH, Kano.

28. Figure : Type of TB and HIV co-infection among study subjects at IDH, Kano

29. Table : Prevalence and Rifampicin resistance among Tuberculosis patients in Kano

30. Plate 1: Band images for Locus MIRU – VNTR locus 424

31. Plate 2: Band images for Locus MIRU – VNTR loci 580 and 1644

32. Figure 9: MIRU-VNTR database for describing MTBC isolates

33. Figure : Distribution and frequency of MTBC genotypes circulating in Kano

34. Two isolates have been described as M. africanum West Africa 1 (ST 330) based on similarity but were also recognized as M. canetti and M. tuberculosis Beijing type respectively based on phylogenic tree analysis

35. Figure : Rifampicin susceptibility among the Cameroon lineages in Kano

36. Genotypic characterization of MTBC strains in Kano has indicated a relatively wide genetic diversity with Cameroon lineage being the most predominant Similarly in Calabar Benjamin et al . (2012) reported the LAM10–CAM clade ( SIT61) and was associated with recent transmission among 25–34 years ( p= 0.019), because 56.3% of the patients harboring this lineage belonged to this age group. However Barbara in 2018 discovered Lineage 4 Cameroon sublineage ( L4.6.2) that represented half of the spoligotyping patterns and M. africanum (L5 and L6) represented one fifth similar to what we reported though we use a more discriminatory genotypic tool (MIRU-VNTR).

37. RECOMMENDATIONS INVESTMENT IN HEALTHCARE IMPROVED STRATEGIC PARTNERSHIPS TO ATTRACT NEW POTENTIAL DONOR ORGANISATIONS INNOVATIVE HEALTH SERVICES AND SOLUTIONS FROM RESEARCHES DONE ON TB HUMAN RESOURCES FOR HEALTH (capacity building to meet challenges of modern healthcare technology ) 5. RETHINKING GLOBAL HEALTH FUNDING FOR THE GOOD OF ALL ( the economic burden of TB treatment and control)

38. REFERENCES Contact the presenter for detailed references Weniger T, Krawczyk J, Supply P, Niemann S, Harmsen , D. MIRU- VNTRplus : a web tool for polyphasic genotyping of Mycobacterium tuberculosis complex bacteria. Nucleic Acids Res 2010, 38 Suppl:W326-331 Allix-Béguec C, Harmsen D, Weniger T, Supply P, Niemann S. Evaluation and user-strategy of MIRU- VNTRplus , a multifunctional database for online analysis of genotyping data and phylogenetic identification of Mycobacterium tuberculosis complex isolates. J Clin Microbiol 2008, 46(8):2692-2699 Kamerbeek J, Schouls L, Kolk A, et al. (1997). Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J Clin Microbiol 35:907-914. http ://www.miru-vntrplus.org

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