Comparison of LCD array and IS6110-PCR with conventional techniques for detection of Mycobacterium bovis isolated from Egyptian cattle and Buffaloes

Rasha Nabil Zahran; Ayman El Behiry; Eman Marzouk; Tamer Askar

doi:10.1016/j.ijmyco.2014.06.002

Comparison of LCD array and IS6110-PCR with conventional techniques for detection of Mycobacterium bovis isolated from Egyptian cattle and Buffaloes

Int J Mycobacteriol. 2014 Sep;3(3):197-204. doi: 10.1016/j.ijmyco.2014.06.002. Epub 2014 Jul 2.

Authors

Rasha Nabil Zahran¹, Ayman El Behiry², Eman Marzouk³, Tamer Askar⁴

Affiliations

¹ (a)Department of Bacteriology, Mycology and Immunology, Faculty of Veterinary Medicine, University of Sadat City, Sadat City, Egypt; (b)Department of Medical Microbiology, Faculty of Applied Medical Science, Taif University, Taraba, Saudi Arabia.
² (a)Department of Bacteriology, Mycology and Immunology, Faculty of Veterinary Medicine, University of Sadat City, Sadat City, Egypt; (c)Department of Public Health (Microbiology), College of Public Health, Qassim University, Saudi Arabia. Electronic address: aymanella2007@yahoo.com.
³ (d)Department of Scientific Unit (Biology), Deanship of Educational Services, Qassim University, Saudi Arabia.
⁴ (e)Consultant of Cardiology in Taif University Clinic, Saudi Arabia.

PMID: 26786488
DOI: 10.1016/j.ijmyco.2014.06.002

Abstract

Bovine tuberculosis is a chronic bacterial and major infectious disease of cattle and buffaloes caused by Mycobacterium bovis. Rapid diagnosis of bovine tuberculosis is considered one of the cornerstones for worldwide control as it permits early epidemiological and therapeutic interventions. Therefore, this study was designed to evaluate conventional techniques (tuberculin test, Ziehl Neelsen staining and culturing) in comparison with proven molecular laboratory techniques (LCD array and IS6110 PCR) for identification of Bovine tuberculosis. A total of 902 Egyptian animals (480 buffaloes and 422 cattle) were examined by tuberculin test, and the positive reactors were slaughtered. Tissue samples were collected for staining as well as culturing. Moreover, LCD array and PCR using IS6110 on DNA extracted from tissue and culture samples were carried out for molecular identification of M. bovis. According to the results, the tuberculin positive cases for cattle and buffaloes were 2.14% (9 cases) and 5.62% (27 cases), respectively. After post-mortem examination, the prevalence of tuberculin positive cases with visible lesions was 88.9% for cattle and 14.8% for buffaloes. Alternatively, these percentages were 11.1% and 85.2% for cattle and buffalo carcasses with non-visible lesions. The percentage of cattle and buffaloes showing positive culture was 88.9% and 62.9%, respectively. This percentage was 69.5% after staining with Ziehl Neelsen. In contrast, LCD array and IS6110 were 100%, confirming the isolation results. In conclusion, LCD array depending on 16S RNA and DNA hybridization with specific probes for detection of M. bovis are rapid, sensitive and labor-saving when combined with IS6110-PCR.

Keywords: Culture; IS6110-PCR; LCD array; Mycobacterium bovis; Tuberculin.