Basic knowledge about MTB Mycobacterium tuberculosis, a small, aerobic, non-motile bacillus. "Gram-positive“ Tuberculosis typically spread through the air when people who have an active TB infection cough, sneeze etc. symptoms The classic symptoms of active TB infection are a chronic cough with blood-tinged sputum, fever, night sweats, and weight loss. Diagnosis Radiology (commonly chest X-rays) microbiological culture. tuberculin skin test (TST) etc

1. Mycobacterium tuberculosis
Presented by
ARCHANA GAUTAM
3rd,semester
BABA SAHEB BHIMRAO AMBEDKER UNIVERSITY
LUCKNOW

2. Abstract
Strain-specific genomic diversity in the Mycobacterium tuberculosis complex (MTBC) is an
important factor in pathogenesis that may affect virulence, transmissibility, host response and
emergence of drug resistance. Several systems have been proposed to classify MTBC strains into
distinct lineages and families. Here, we investigate single-nucleotide polymorphisms (SNPs) as
robust (stable) markers of genetic variation for phylogenetic analysis. We identify ~92k SNP
across a global collection of 1,601 genomes. The SNP-based phylogeny is consistent with the
gold-standard regions of difference (RD) classification system. Of the ~7k strain-specific SNPs
identified, 62 markers are proposed to discriminate known circulating strains. This SNP-based
barcode is the first to cover all main lineages, and classifies a greater number of sublineages
than current alternatives. It may be used to classify clinical isolates to evaluate tools to control
the disease, including therapeutics and vaccines whose effectiveness may vary by strain type.

3. Introduction
Dr. Robert Koch discovered
the
tuberculosis bacillus. (1882)
He received the Nobel Prize in
physiology or medicine in 1905 for this
discovery
After 100 years later
The bacillus causing tuberculosis,
airborne disease, M.tuberculosis
was identified and described on 24
March 1882 by Robert Koch .

4. Mycobacterium tuberculosis(MTB)
Basic knowledge about MTB
• Mycobacterium tuberculosis, a small, aerobic, non-motile bacillus.
• "Gram-positive“
• Tuberculosis typically spread through the air when people who have
an active TB infection cough, sneeze etc.
symptoms
• The classic symptoms of active TB infection are a chronic cough
with blood-tinged sputum, fever, night sweats, and weight loss.
Diagnosis
• Radiology (commonly chest X-rays)
• microbiological culture.
• tuberculin skin test (TST) etc.

5. M.tuberculosis cont......
M. tuberculosis under
microscope M. tuberculosis under SEM
Kingdom : Bacteria
Phylum: Actinobacteria
Order: Actinomycetales
Suborder:Corynebacterineae
Family: Mycobacteriaceae
Genus: Mycobacterium
Species: M.tuberculosis
M. tuberculosis
Bacterial colonies

6. Tb affects many parts
of body but mainly
divided into two parts
i.e. Pulmonary and.
Extra-pulmonary

7. TB Drug resistance MDR

8. Cont....

9. Cont.....

10. Barcode
Single nucleotide
polymorphism
(SNP)
According
to
research
paper

11. Why SNPs ? ? ?
 The selection of suitable SNPs is crucial for the success of a typing assay.. The need
for a suitable sequence .
 SNPs for were chosen using following criteria:
 SNP is specific for the lineage of interest
 SNP is not in a region where a genomic deletion has been described (to ensure
binding of oligonucleotides and subsequent allele-calls for all strains)
 No other SNP less than 50 bp up- or downstream (to avoid mismatches in
oligonucleotide annealing)
 SNP is not a transition (i.e. no nucleotide change A>G, G>A, C>T or T>C; lower
unspecific signal because no alternate base pairing possible)
 SNP is in a coding sequence (deletions or insertions less likely)
 SNP is in an essential gene (deletions less likely)
 SNP change is synonymous (no amino acid change; therefore lower selective
pressure acting on SNP compared to non-synonymous SNP)

12. SNPs
(molecular marker)
• David Stucki et al (2012) to assess genotype-phenotype associations,
phylogenetically robust molecular markers and appropriate genotyping tools are
required.

14. Treatment success
has recently
exceeded the
global target of
85% Case
Detection in DOTS
areas has recently
attained global
target of 70%
TB
in
INDIA
Drug resistance problem in India and globally
was worsening

15. INDIA is the highest TB burden country accounting for more
than one-fifth of the global incidence
Indonesia
6%
Nigeria
5%
Other countries
20%
Other 13 HBCs
16% China
14%
South Africa
5%
Bangladesh
4%
Ethiopia
3%
Pakistan
3%
Phillipines
3%
India
21%
Source: WHO Geneva; WHO Report 2014: Global Tuberculosis Control; Surveillance, Planning and Financing
Global annual incidence = 9.4 million
India annual incidence = 1.96 million

16. MDR-TB cases in the hospital
The number of
(MDR)-TB cases
in the country
had increased
five-fold
between
2011and2014.
Studies show
one-third of the
MDR-TB cases
are resistant to
fluoroquinolones
, which are
critical for MDR-
TB treatment.
Deshakalyan Chowdhury/Agence France-Presse/Getty Images
Tuberculosis patients at a hospital in Kolkata, April 07, 2008

17. Problem of TB in India
• Estimated incidence
– 1.96 million new cases annually
– 0.8 million new smear positive cases annually
– 75 new smear positive PTB cases/1lakh population per year
• Estimated prevalence of TB disease
– 3.8 million bacillary cases in 2000
– 1.7 million new smear positive cases in 2000
• Estimated mortality
– 330,000 deaths due to TB each year
– Over 1000 deaths a day
– 2 deaths every 3 minutes

18. Problem of TB in India (contd)
• Prevalence of TB infection
– 40% (~400m) infected with M. tuberculosis (with a 10% lifetime risk
of TB disease in the absence of HIV)
• Estimated Multi-drug resistant TB
– < 3% in new cases
– 12% in re-treatment cases
• TB-HIV
– ~2.31 million people living with HIV.
– 10-15% annual risk (60% lifetime risk) of developing active
TB disease.
– Estimated ~ 5% of TB patients are HIV infected

19. This Wednesday, the WHO declared drug-resistant TB a public health crisis, saying the airborne disease is rising faster than
countries can treat it. The WHO also said many cases are still going unreported
Evolution of TB Control in India
• 1950s-60s Important TB research at TRC and NTI
• 1962 National TB Programme (NTP)
• 1992 Programme Review
• only 30% of patients diagnosed;
• of these, only 30% treated successfully
• 1993 RNTCP pilot began
• 1998 RNTCP scale-up
• 2001 450 million population covered
• 2004 >80% of country covered
• 2006-till today Entire country covered by RNTCP

20. 0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
110%
120%
2000 2001 2002 2003 2004 2005 2006 2007 2008 2009
Annualised New S+ve CDR Success rate
•Population projected from 2011 census
•Estimated no. of NSP cases - 75/100,000 population per year (based on recent report)
Annualized New Smear-Positive(NSP) Case Detection Rate (CDR)
and Treatment Success Rate(TSR) in DOTS Areas, India, 2002-2014*
2002 2003 2004 2005 2007 2008 2009 2010 2011-14
% TSR

22. Articles/News on TB in INDIA• Drug-resistant TB cases on rise in north India, reveals
PGI research Express News Service : Chandigarh, Thu May 09
2013, 02:33 hrs
• Study finds doctors also adding to burden of drug-
resistant TB cases Times Of India Jul 19, 2014, 01.13 AM
• NEW DELHI: In the wake of multi-drug resistant (MDR)-
TB cases increasing in India, Union Health Minister Harsh
Vardhan on 6th,september 2014 launched India's first
National Anti-Tuberculosis Drug Resistance Survey, 2014-
2015. (NDTV REPORT)
• "It often happens that patients who have TB disease
stop taking the drugs soon after they start feeling
better. They can become sick again as if the drugs are
not taken correctly, the TB bacteria may still remain
alive and the person may become resistant to those
drugs and will have to then given second-line drugs,
“(NDTV REPORT)
• In December, India’s director general of health services,
Dr. Jagdish Prasad told The Wall Street Journal that India
planned to provide free drugs to patients being treated by
private doctors so that the government could make sure all
patients are properly tested and have an accurate
assessment of disease burden (Wall Street Journal )

23. Discussion
• In conclusion, using the whole-genome sequences of over 1,601 MTBC
isolates, we characterize a high-resolution map of polymorphisms
consisting of more than ninety thousand SNPs. This genomic variation is
used to infer phylogenetic relationships both inter- and intra-lineage to an
unprecedented level of resolution, and lead to the development of an
extendable nomenclature for sublineages. We identify a panel of 62 robust
SNP markers (of 413 suitable alternatives) that can be used to construct
high resolution and reproducible phylogenies, which can be incorporated in
diagnostic assays and assess genotype–phenotype associations. Future
work should focus on other types of lineage-specific polymorphisms (for
example, insertions, deletions and large structural variants), which are less
common than SNPs, but may have major functional consequences. The
proposed system has the flexibility to incorporate novel strain types should
they be reported. Incorporating anti-tubercular drug-resistance loci will
further enhance the usefulness of the barcode as an important tool for
tuberculosis control and elimination activities worldwide.

24. Result
• A genomic analysis was performed on whole-genome sequences of 1,601
MTBC isolates from eleven independent sequencing studies from different
areas of the world.
• The technique could identify super-spreaders and predict the existence of
undiagnosed cases, potentially leading to early treatment of infectious
patients and their contacts.
• It may be used to classify clinical isolates to evaluate tools to control the
disease.
• In future decreased the rate of multi drug resistance (MDR) TB.

25. References
Research paper’s references
1.David Stucki, Bijaya Malla, Simon Hostettler, Thembela Huna,et al(2012) Two
New Rapid SNP-Typing Methods for Classifying Mycobacterium tuberculosis
Complex into the Main Phylogenetic Lineages.
2.Gutacker MM, Mathema B, Soini H, Shashkina E, Kreiswirth BN, et al. (2006)
Single-nucleotide polymorphism-based population genetic analysis of
Mycobacterium tuberculosis strains from 4 geographic sites. J Infect Dis 193:
121–128. doi:10.1086/498574.
3.Wang L, Luhm R, Lei M (2007) SNP and mutation analysis. Adv Exp Med Biol
593: 105–116. doi:10.1007/978-0-387-39978-2_11.
Book’s references
1.Brock biology of microorganism Madigan et al (2011) 13th,edition
2.Plant biotechnology the genetic manipulation of plants Slater et al (2012)
2nd,edition
3.Prescott 7th, edition

26. THANK YOU
CARELESS
SPITTING, COUGHING,SNEEZING
SPREAD TUBERCULOSIS