Title Evaluation of Three Molecular Methods of Repetitive Element Loci for Differentiation of Mycobacterium avium subsp. paratuberculosis (MAP)
Author Amr El-Sayed1,2, Abdulwahed Ahmed Hassan1,3*, Saleh Natour1, Amir Abdulmawjood4, Michael Bulte4, Wilfried Wolter5, and Michael Zschock1
Address 1Landesbetrieb Hessisches Landeslabor, Schubertstrasse 60, Haus 13. D-35392, Giessen, Germany, 2Institute of Internal Medicine and Animal Infectious Disease, Faculty of Veterinary Medicine, Cairo University, 12211 Giza, Cairo, Egypt, 3GD Animal Health Service, Department of Bacteriology and Parasitology, Postbus 9, 7400 AA Deventer, The Netherlands, 4Institut f? Tier?ztliche Nahrungsmittelkunde, Justus-Liebig-Universit?, Frankfurterstr. 92, Giessen, Germany, 5Regierungspr?idum Giessen, Dez. 51.2, Schanzenfeldstrasse 8, 35578 Wetzlar, Germany
Bibliography Journal of Microbiology, 47(3),253-259, 2009,
Key Words Mycobacterium avium subsp. paratuberculosis (MAP), short sequence repeats (SSR), mycobacterial interspersed repetitive units (MIRU), variable number tandem repeats (VNTR), Johne''s disease (JD)
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.