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Evaluation of three molecular methods of repetitive element loci for differentiation of Mycobacterium avium subsp. paratuberculosis (MAP)

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Abstract

The aim of the present study is to evaluate the efficiency of three methods to determine the molecular diversity of 34 Mycobacterium avium subsp. paratuberculosis (MAP) strains isolated from 17 cattle herds. The applied methods included the analysis of sequence polymorphism of the mononucleotide (G1 and G2) and trinucleotide sequences (GGT) of the Short Sequence Repeats (SSR) and the determination of size polymorphism of 9 different Mycobacterial Interspersed Repetitive Units (MIRU) and 6 Variable Number Tandem Repeats (VNTR). Sequence analysis of SSR of 34 isolates showed 4, 6, and 2 alleles of G1, G2, and GGT repeats, respectively. The amplification of the investigated 9 MIRU units revealed only two discriminatory genotyping systems (MIRU2 and MIRU3). Out of 6 VNTR PCR differentiation methods, only one method could be recommended for genotyping purposes. The profile 7g-12g-4ggt-II-b-2 of the combination systems G1-G2-GGT-MIRU2-MIRU3-VNTR1658 dominates among the examined isolates and was detected in 14.7% of the isolates. The use of certain repetitive loci of SSR, MIRU, and VNTR techniques in this study showed greater potential than others for the characterization of MAP isolates. The recommended loci can be used for the epidemiological tracing of MAP field strains and to determine the relationships between isolates in different herds.

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Authors and Affiliations

  1. Landesbetrieb Hessisches Landeslabor, Schubertstrasse 60, Haus 13, D-35392, Giessen, Germany

    Amr El-Sayed, Abdulwahed Ahmed Hassan, Saleh Natour & Michael Zschöck

  2. Department of Bacteriology and Parasitology, GD Animal Health Service, Postbus 9, 7400 AA, Deventer, The Netherlands

    Abdulwahed Ahmed Hassan

  3. Institut für Tierärztliche Nahrungsmittelkunde, Justus-Liebig-Universität, Frankfurterstr. 9 2, Giessen, Germany

    Amir Abdulmawjood & Michael Bülte

  4. Regierungspräsidum Giessen, Dez. 51.2, Schanzenfeldstrasse 8, 35578, Wetzlar, Germany

    Wilfried Wolter

  5. Institute of Internal Medicine and Animal Infectious Disease, Faculty of Veterinary Medicine, Cairo University, 12211, Giza, Cairo, Egypt

    Amr El-Sayed

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  1. Amr El-Sayed
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  2. Abdulwahed Ahmed Hassan
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  3. Saleh Natour
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  4. Amir Abdulmawjood
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  5. Michael Bülte
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  6. Wilfried Wolter
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  7. Michael Zschöck
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Correspondence to Abdulwahed Ahmed Hassan.

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El-Sayed, A., Hassan, A.A., Natour, S. et al. Evaluation of three molecular methods of repetitive element loci for differentiation of Mycobacterium avium subsp. paratuberculosis (MAP). J Microbiol. 47, 253–259 (2009). https://doi.org/10.1007/s12275-008-0257-1

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  • DOI: https://doi.org/10.1007/s12275-008-0257-1

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