BY AMIT TYAGI Under the supervision of Dr. Yogesh KumarSingh

Tuberculosis: A disease caused by tiny germs that enter in lungs when someone breath them in.

Mycobacterium tuberculosis is a rod shaped, non motile, acid fast bacterium

Symptoms: Cough Chills Afternoon fever Weight loss Night sweat

In 2008 China (1.3 million), South Africa (0.47 million), Nigeria (0.45 million) and Indonesia (0.43million)

Each year, there are 8 million people diagnosed with TB and 2 million deaths worldwide.

India is 17th among 22 High Burden Countries. India (1.98 million).

It is a major socio-economic burden in 5–16 % of cases of infertility among Indian women.

TB Global StatusTB Global Status

There are several methods/techniques are used worldwide for the detection of TB such as-

Bactec 460 TB system, ZN staining, LJ medium, PCR,Skin Test etc.

Among all these technique BACTEC 460 & PCR are used commonly in medical science and research labs widely indicating that these are more specific than others.

When the BACTEC 12B vial is inoculated, mycobacterium, if present, utilize the 14C labeled substrate (fatty acid) and release 14CO2.

BACTEC instrument measures quantitatively the radioactivity on a scale ranging from 0-999, as GI.

BACTEC 460 is Radiometric method.

The BACTEC system gives early culture results with differentiation of M. tuberculosis from (MOTT).

Using the same system drug

susceptibility test can also be performed for

all the anti-tuberculosis drugs

when sufficient GI is observed.

Tests are performed in

vitro, and measure the

growth response of an isolated organism to a

particular drug or drugs.

When an Anti tuberculosis drug is added to the medium, suppression of growth occurs if the test organism is susceptible.

Sample

Decontamination

Proceeds to Culture

Subculture

Cultured in BACTEC460

Inoculated sample in 12B vials

Compare of growth on the basis of growth index

Growth index reading

Drugs Susceptibility

Differentiation of MTB from MOTT with the help of NAP test

Detection of TB with Detection of TB with BACTEC 460BACTEC 460

Sample

DNA isolation

Isolated DNA Master mix preparation

Amplified DNA segments

Compare bands in UV light

DNA Amplification in Thermo cycler

Gel electrophoresis

The amplified sequence can then be detected by Agarose gel electrophoresis.

Comparative study of BACTEC460 and PCR for the detection of T.B.

Comparative study of positivity ratio in various samples.

Comparative study of gender biasness of T.B. infections.

Comparative study of positivity ratio in various age groups.

Study of drug sensitivity with positive samples.

Collection center • Sample were collected from various reputed

hospitals & pathology labs like..

Sample collection

Total no. of samples detected by BACTEC 460 and PCR

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BACTEC POSITIVITY 66.66% 29.16% 16.66% 12.5% 0%PCR POSITIVITY 66.66% 33.33% 29.16% 50% 100%

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BACTEC POSITIVITY 0% 17.59% 19.44% 42.85% 0%PCR POSITIVITY 25% 37.03% 27.77% 28.57% 0%

Comparison drugs sensitivity & resistant in total sample

GI (control) > GI (drug) – “susceptible” 

GI (control) < GI (drug) – “resistant” 

GI (control) = GI (drug) – “borderline”

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Pie chart for drug resistance

PCR proved better diagnostic method then BACTEC 460 TB radiometric system

BACTEC 460 TB radiometric system resulted in the total positivity values of 19.91% and PCR, the total sensitivity value of 35.18%.

Males are more susceptible for tuberculosis then Females in the NCR region.

In given data, younger (13) as well as older (83) age patients can be affected with tuberculosis,

Females of age group 20-40 and Males of age group 80-100 are more susceptible for tuberculosis.

Data suggest that maximum infected patients are sensitive towards Pyrazinamide & Streptomycin & resistant for Ethambutol