1 / 38

Molecular Typing of Mycobacterium spp. Isolates from Tuberculosis Patients

This study aims to characterize and identify Mycobacterial species isolated from TB patients using conventional and molecular techniques. PCR is an important tool in diagnostic medicine for rapid and accurate identification. The use of molecular techniques before treatment initiation can provide valuable information on the epidemiology and treatment of TB.

goble
Download Presentation

Molecular Typing of Mycobacterium spp. Isolates from Tuberculosis Patients

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Molecular typing of Mycobacterium spp. isolates fromtuberculosis patientsDr. Anas Al-MahbashiAssistant Professor of Molecular Biology-.Physiology Department –Faculty of Medicine – Al-Kalamoon University- 2019.

  2. (تحديد الأنماط الجزيئية للمستفردات من أنواع المتفطرات لدى مرضى السل)د. أنس احمد المحبشيدكتوراة في الأحياء الجزيئيه والأحياء الدقيقه الطبيهأستاذ مساعد- كليه الطب البشري – جامعه القلمون الخاصه – سوريا2019.

  3. Tuberculosis (TB) is a disease of major public health concern worldwide.It is a bacterial infectious disease that is considered the second most important cause of death due to infectious agent

  4. # one-third of the world’s population is infected with latent TB 5%–10% of this population will develop active stages of the disease during their life time (Reactivation)

  5. infection can occur via inhalation of droplet particles aerosolized from persons infected with Mycobacteriumtuberculosis or by consumption of milk infected with bovine M. bovis

  6. -Overcrowding & a rapidly increasing population, -poor quality health services -Poor sanitation -low income & poor economic status -substandard housing with poor ventilation are the most important factors responsible for the high incidence of TB in developing countries

  7. in developing countries although the disease has declined but ; the disease re-emerged again due to:- • increasing number of HIV individuals • Homeless • Prison inmates • Illegal Immigration

  8. In Syria The absolute number of TB cases is not known, but the total cases notified of Tb infection 2 845 cases (WHO, 2017).

  9. Geographical Distribution o TB

  10. Although the disease mainly infect the Lung (Pulmonary TB)But The disease may involve all Body Organs (Extra-Pulmonary TB).

  11. The Mycobacterium genus comprises more than 70 different species and is distributed worldwide. Mycobacteria that cause TB. in mammals form the Mycobacterium tuberculosis complex (MTC) which include:- M. tuberculosis, M. afrcanum, M.bovis M. micrott and M. canett.

  12. Other forms of Mycobacteria that are termed Mycobacteria other than Mycobacterium tuberculosis (MOTT).

  13. Treatment of Tb is not like other bacterial infections. It is complicated & need long time course.It lasts for 6-8 months & requires combination of 4-5 different antibiotics drugs.

  14. Unfortunately in the recent years the appearance of Multi drugs resistant (MDR) strains hampered the tuberculosis control efforts & increase the size of problem in the world.

  15. The objective :- To characterize & identify Mycobacterial Species isolated from TB patients using conventional and Molecular technique.

  16. PCR is one of the most important and powerful technique, Particularly in Diagnostic Medicine . • In 1985, Kary Mullis invented a process he called PCR, which solved a core problem in genetics: How to make copies of a strand of DNA you are interested in.

  17. Sample collection (120 ) Sputum samples were collected from Yemeni Patients with suspected pulmonary TB. -Samples collected from National Tuberculosis Center in Sana’a city - Yemen

  18. -Staining processing -Cultivation Of Bacteria In specialist Media -Antibiotics sensitivity test. -DNA extraction

  19. DNA extraction, preservation

  20. Molecular Technique was done In the Institute of Endemic diseases –Faculty Of Medicine –Khartoum University – Sudan.Polymerase Chain Reaction ( PCR-RFLP).

  21. Primers specific for the rpoB gene, (rpoB DNA, 342–360 base pairs) was the target region for amplification and identification of Mycobacterium spp

  22. mycobacterial DNA was amplified successfully using the relevant primers and the size of DNA was 360 bp compared withthe molecular weight marker (100bp)

  23. Following amplification of the rpoBgene. the PCR- product was subjected todigestion by restriction enzymes Restriction fragment length polymorphism (RFLP)

  24. DNA sequencing Analysis

  25. DNA sequencing of MTC is different from MOTT.Interestingly theMOTT isolates were resistant to isoniazid, rifampicin and streptomycinand sensitive only to Ethambutol.

  26. PCR technique yield improvement in methodology in the mycobacteriology laboratory, It showed more rapid and accurate identification of Myccobacterium from clinical specimens.

  27. It is important in TB treatment to determine if the infection is due to MTC or MOTT because many of these medication have side effects, and need continuous monitoring. Treatment may be necessary for as long as two years in some cases.

  28. we recommend the use of molecular techniques for identification of the Mycobacteriumspp. before initiation of treatment

  29. Molecular technique is a good tools in diagnosis of TB disease:- -Rapid & accurate identification -More specific & more sensitive Gave knowledge on the epidemiology & Treatment manner of disease.

  30. Thanks You

More Related