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abstract
Molecular genotyping and epidemiology of Mycobacterium bovis strains obtained from cattle in Khorasan province, Iran: 2010-2012
Ahmadi M.1, Mosavari N.2, Tadayon K.2, Soleymani K.2, Dashtipour S.2, Arefpajoohi R.2, Keshavarz R.2, Mohammad Taheri M.2
1 Islamic Azad University, Damghan, IRAN
2Razi Vaccine and Serum Research Institute, Karaj, IRAN

Key words: Mycobacterium bovis, PGRS-RFLP, DR-RFLP

Introduction: Mycobacterium bovis is a pathogen of significant importance in livestock and a wide range of wild animal species worldwide. It is also known to cause tuberculosis disease in humans, a fact which has raised renewed concerns regarding the zoonotic risk for humans, especially those living at the animal-human interface. Molecular typing methods such as restriction fragment length polymorphism (RFLP) analysis have been used to identify the source of infection into cattle herds and to trace the transmission of bovine tuberculosis that is essential to control and eradicate of this disease.
Materials and methods: In this study, Restriction fragment length polymorphism (RFLP) genotyping was employed to analyze the population genetics of Mycobacterium bovis in Khorasan province of Iran. Sixty one isolates collected from slaughtered tuberculosis-suspect cattle were subjected to RFLP analysis with probes of the polymorphic GC-rich sequence (PGRS) and the direct repeat sequence (DR) using DNA digested with PvuII.

Results: Genotyping of the isolates by PGRS-restriction fragment length polymorphism (RFLP) and DR-RFLP with Pvu II displayed 2 and 3 genotypes were identified by PGRS and DR respectively. PGRS-RFLP and DR-RFLP methods detected homogeneous population in which only a few isolates had variant genotypes which was consistent with other previous studies in Iran.

Conclusion: Hopefully, in the future we will have new and improved techniques for typing M. bovis. It is conceivable that a lab-on-a-chip approach will be capable of not only detecting M. bovis from a clinical sample but also typing the pathogen at the same time.

Islamic Azad University
Damghan Branch
Faculty of

A Thesis Submitted in Partial Fulfillment of the Requirments For The Degree of M.sc in

Title
Molecular genotyping and epidemiology of Mycobacterium bovis strains obtained from cattle in Khorasan province, Iran: 2010-2012

Supervisor
Dr Nader Mosavari

Advisor
Dr Kyvan Tadayon

By
Mahboobeh Ahmadi

February 2014

1Mycobacterium
2 Huyen, Kremer
3 Singh , Verma
4 David et al
2 Restriction Fragment Length Polymorphisms (RFLP)
6Parreiras et al
4 Polymophic GC-rich Repeatative Sequences (PGRS(
5 Direct repeat (DR) sequences
9 Mi Hee et al
10 Muayad et al
11 Limeschenko et al
12 Asiimwe, Med
13Rastogi et al
1 Gutierrez et al
15 Durr et al
16M. africanum
17M. microti
18M. simiae
19M. avium
20M. phlei
21M. smegmatis
22M. avium- M.intracellular- M.scrofulaceum (MAIS)
23M.intracellular
24M.scrpfulaceum
25DNA-hybridisition
26 Arunee Thong-On et al
27Mycobacteriaother than the M. tuberculosis complex
28M.lepra
29 OIE
30 Sezavar
31 Cole
32Kamerbeek et al
33 Hlokwa et al
34Runyon
35 Diamond
36Michel
37 Brosch et al
1Amplification
39Polymerase chain reaction
40 Alonso et al
41Primer
42Freezing and thawing
43Cross contamination
44Insertion sequence
45Beer et al
46Multiplex PCR
47Goodfellow &Wayne
48 Neimann et al
49 Shinnick & Good
50 Bull et al.
51 Heifets
52 Mc fadden
53 Brand
54 Boer et al
55 Insertion sequense
56 Costello et al & Beenhouwer et al
57 Spoligotyping
58Variable number tandem repeat
59Multiplex PCR
60 Restriction Fragment Length Polymorphism
61 Southern blotting
62 Restriction Endonuclease Analysis
1 Restriction endonucleases
2 Transposons
3 Insertion sequences
4 Jumping genes
67 Hooper et al
6 Van Soolingen

7 Direct repeat (DR) sequences

8 Spoligotyping

1 Southern Blotting
2 Chromogen digoxigenin
73 Haas et al
74 Yokoyama
2 hot spots
76 Warren
77 MC Hugh
78 Buer
79 Borgdorff
80 Skuce
6 Polymorphic GC-rich Repeatative Sequences(PGRS)
82 Ross
83 Costello
84 Z.H.Yang
59 Direct repeat restriction fragment length polymorphism
86 Groenen
87 Zumarrage
2 Spoligotyping
PCR 3
90 Homolka
Direct Repeat 5
6 Direct Variable Repeat

93 Homolka
94 Gutierrez 2006;Milan 2004
3 Variable Number Tandem Repeat
96 Frothingham
Tandem Repeats Exact 1

2 Insertion
3 Deletion
4 Major Polymorphic Tandem Repeats

101 Fillol
102 Benjamin Marten
103 Austin Flint
104 Jing
105 Boer
5 Rivolta
6 Maffuci
1 Smith

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