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BY AMIT TYAGI Under the supervision of Dr. Yogesh KumarSingh

Comparative Study of BACTEC 460 System and PCR For The Detection of Tuberculosis AND Drug Sensitivity Test By BACTEC 460 TB System. BY AMIT TYAGI Under the supervision of Dr. Yogesh KumarSingh Sr. Scientist AUROPROBE LABORATRIES MURADNAGAR, U.P.

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BY AMIT TYAGI Under the supervision of Dr. Yogesh KumarSingh

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  1. Comparative Study of BACTEC 460 System and PCR For The Detection of Tuberculosis AND Drug Sensitivity Test By BACTEC 460 TB System BY AMIT TYAGI Under the supervision of Dr. YogeshKumarSingh Sr. Scientist AUROPROBE LABORATRIES MURADNAGAR, U.P.

  2. Introduction 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

  3. TB Global Status Each year, there are 8 million people diagnosed with TB and 2 million deaths worldwide. In 2008 China (1.3 million), South Africa (0.47 million), Nigeria (0.45 million) and Indonesia (0.43million) India is 17th among 22 High Burden Countries. India (1.98 million). It is a major socio-economic burdenin 5–16% of cases of infertility among Indian women.

  4. DIAGNOSIS OF TB • 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.

  5. BACTEC 460 TB System BACTEC 460 is Radiometric method. 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. The BACTEC system gives early culture results with differentiation of M. tuberculosis from (MOTT).

  6. Tests are performed in vitro, and measure the growth response of an isolated organism to a particular drug or drugs. Using the same system drug susceptibility test can also be performed for all the anti-tuberculosis drugs when sufficient GI is observed. When an Anti tuberculosis drug is added to the medium, suppression of growth occurs if the test organism is susceptible.

  7. Detection of TB with BACTEC 460 Sample Decontamination Proceeds to Culture Inoculated sample in 12B vials Compare of growth on the basis of growth index Cultured in BACTEC460 Subculture Differentiation of MTB from MOTT with the help of NAP test Drugs Susceptibility Growth index reading

  8. Detection of TB with P C R Sample DNA isolation Master mix preparation Isolated DNA DNA Amplification in Thermo cycler Amplified DNA segments Gel electrophoresis Compare bands in UV light The amplified sequence can then be detected by Agarose gel electrophoresis.

  9. Objectives 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.

  10. Sample collection Collection center • Sample were collected from various reputed hospitals & pathology labs like.. • AIIMS, • GTB, • Metro Noida, • Yashoda K & N, • Max Noida, • Sarvodaya Hospital, • Dr. Lal Path Lab, • Dr. Khanna Lab, • Phoenix Hospital, • Batra Hospital, • Metro Hosp. Lajpatnagar, • Dr. J. Bansal, • Fortis Escorts Hospital etc

  11. Comparison Between BACTEC & PCR Total no. of samples detected by BACTEC 460 and PCR Number of samples

  12. Comparison in Female Sample Number of samples

  13. Comparison in Male Sample Number of samples

  14. Comparison in Different Age Group of Male Number of samples BACTEC POSITIVITY 66.66% 29.16% 16.66% 12.5% 0% PCR POSITIVITY 66.66% 33.33% 29.16% 50% 100%

  15. Comparison in Different Age Group of Female Number of samples BACTEC POSITIVITY 0% 17.59% 19.44% 42.85% 0% PCR POSITIVITY 25% 37.03% 27.77% 28.57% 0%

  16. GI (control) > GI (drug) – “susceptible”  • GI (control) < GI (drug) – “resistant”  • GI (control) = GI (drug) – “borderline” Drugs Sensitivity Test Number of samples Comparison drugs sensitivity & resistant in total sample

  17. Percentage of Drug Sensitivity & Resistant

  18. Drug resistance In Males and Females Pie chart for drug resistance

  19. Conclusion 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.

  20. 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

  21. Thank you

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