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DNA fingerprinting of
M.tuberculosis isolates from
Chakkipath region of Agra
by using IS6110 probe
Under supervision of :-
Submitted By:-
Dr. D.S Chauhan
Lata Kumari
Scientist D
INTRODUCTIO
N
Tuberculosis is an ancient and
infectious disease caused by various
strains of Mycobacterium tuberculosis.
In 1882 Robert koch isolate and
identified Mycobacterium tuberculosis
& hence known as koch bacillus.
These are slim gram positive acid
fast bacilli (0.4µm x 2-10µm).
They are nonmotile obligate
aerobes and non sporing .
One third of world population has
been infected with M.tuberculosis, and
new infections occur at a rate of one
per second on a global scale.
According to WHO annual report 2012
there were an estimate of 8.7 million
incident cases of TB globally.
In India Uttar Pradesh has highest
incidence rate of about 175507 cases
per years.
There are several diagnostic methods
are available for the epidemiological
studies of mycobacterium strains but
OBJECTIVES OF THE
STUDY
•The main objective of study “DNA
fingerprinting of Mycobacterium
tuberculosis isolates from
Chakkipath region of Agra by
using IS6110 probe”.
•To observe the transmission of the
disease by using this marker .
IS6110 – Restriction
Fragment Length
Polymorphism
The term RFLP refers to the differences in
homologous DNA sequences detected by
presence of fragments with varying lengths after
digestion of DNA .
In RFLP analysis, the DNA sample is broken into
pieces (digested) by restriction enzymes and the
resulting restriction fragments are separated
according to their lengths by agarose gel
electrophoresis.
It uses the insertion sequence IS6110 which is
present in most of the strains of tubercle bacilli
(copy no. 0-25) as a probe to enable the strain
IS6110 RFLP has been used in many
epidemiological studies to detect
outbreaks and to track TB transmission in
large population, to uncover laboratory
cross-contamination and to differentiate
relapse caused by endogenous
reactivation from re- infection by strain .
D DNA Isolation
Restriction of DNA
Electrophoresis
Southern blotting
Prehybridization
Detection
Analysis of results
RESULT
1 2 3 4 5 6 7 8 9 10 11 M
S.No Lane No. Isolate No. Copy No.
1 Lane No. 1 PM 14 9
2 Lane No. 2 PM 16 10
3 Lane No. 3 PM 19 6
4 Lane No. 4 PM 30 9
5 Lane No. 5 PM 34 10
6 Lane No. 6 PM 37 11
7 Lane No. 7 PM 43 16
8 Lane No. 8 PM 57 12
9 Lane No. 9 PM 58 4
10 Lane No. 10 PM 59 14
11 Lane No. 11 PM 61 0
12 Lane No 12 Mol. Marker