Detection of Mycobacterium paratuberculosis by Nested PCR in dairy cattles suspected to john’s disease

Subject Areas : Diagnostic Microbiology

Abbas Doosti ¹ (Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran)
Saadat Moshkelani ² (Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran)

Received: 2009-04-14 Accepted : 2009-05-15 Published : 2009-04-01

Keywords: Mycobacterium paratuberculosis, Nested-PCR, John’s disease, Dairy cattles,

Abstract :

Backgorund and Objectives: Johne’s disease is a chronic intestinal disease of domestic and wild ruminants that is caused by Mycobacterium paratuberculosis. So that Mycobacterium paratuberculosis is a widespread zoonotic disease, necessary to use prevention and control methods to eliminate this agent matter needs the attention of corresponding managers. The aim of this study is base on use of Nested PCR as an accurate and rapid method to detection of Mycobacterium paratuberculosis bovine feces. Material and Methods: Fecal samples from 120 dairy cattle were collected and DNA extraction was performed from the samples. Then, samples were evaluated with specific primers for the Mycobacterium paratuberculosis-specific IS900 gene by Nested-PCR assay. Finally, PCR products were electrophoresis on agarose gel. Results: In total, from 120 samples were enrolled in this study, 22 specimen (18.33%) were found positive on the basis of Nested-PCR analysis. Conclusion: Nested PCR technique is considered as a fast, reliable and affordable assay to detect Mycobacterium paratuberculosis.