ISSN: 2375-3005
American Journal of Microbiology and Biotechnology  
Manuscript Information
 
 
Comparative Study Between Traditional Methods and Molecular Methods in Diagnosis of Bovine Tuberculosis
American Journal of Microbiology and Biotechnology
Vol.4 , No. 6, Publication Date: Oct. 13, 2017, Page: 83-90
237 Views Since October 13, 2017, 48 Downloads Since Oct. 13, 2017
 
 
Authors
 
[1]    

Wessam Youssef, Biotechnology Department, Animal Health Research Institute, Giza, Egypt.

[2]    

Khaled Al-Amry, Microbiology Department, Cairo University, Giza, Egypt.

[3]    

Nashwa Helmy, Biotechnology Department, Animal Health Research Institute, Giza, Egypt.

[4]    

Salah El Deen Abdel Karim, Microbiology Department, Cairo University, Giza, Egypt.

[5]    

Sherif Marouf, Microbiology Department, Cairo University, Giza, Egypt.

 
Abstract
 

Mycobacterium bovis is the major causative agent of bovine tuberculosis (BTB) and part of the Mycobacterium tuberculosis complex (MTBC). BTB has an impact on the national and international economy, affects the ecosystem via transmission to wildlife and is of public health concern due to its zoonotic potential. Although still present in some industrialized countries, BTB today mostly affects developing countries lacking the resources to apply expensive test and slaughter schemes. Tuberculosis (TB) remains a global health problem despite near eradication in some developed countries. This study was conducted from early winter of 2015 to winter of 2017 to compare between conventional and molecular techniques for detection of Mycobacterium bovis (M. bovis) in Egypt. A total of 49 specimens were collected from four major abattoirs (El-Basateen- El-Monieb- Beni-Suef- Al-fayoum) to be analyzed bacteriologically and biochemically for: isolation, identification and confirmation of M. bovis with molecular methods. Only 19 isolates were found to be positive slow-growers Mycobacterium species by conventional cultivation method on solid medium (LJ medium and Stone brink) and identified biochemically to 17 M. bovis isolates and 2 isolates M. tuberculosis. Genotyping detection of Mycobacterium tuberculosis complex by amplification of ext-RD9 region by real-time PCR was carried out on positive cultures and directly on specimens. Out of 49 DNA templates extracted directly from specimens, 31 specimens were confirmed to be infected by Mycobacterium tuberculosis complex by amplification of ext-RD9 region by real-time PCR. This study reports the development and evaluation of a single-tube, two-targets, real-time PCR assay which can differentiate between M. bovis and M. tuberculosis. The multiplex real-time PCR target RD1 and RD4 using 2 sets of primers-probes. A 31 MTBC positive DNA from clinical specimens were identified by this assay as, 27 M. bovis isolates and 4 M. tuberculosis isolates. A 19 positive cultures were confirmed to be Mycobacterium tuberculosis complex were identified by this assay as, 17 M. bovis isolates and 2 M. tuberculosis isolates.


Keywords
 

M. bovis, M. tuberculosis, Real-Time PCR, Multiplex Real-Time PCR


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