Evaluation of the epidemiological relevance of variable-number tandem-repeat genotyping of Mycobacterium bovis and comparison of the method with IS6110 restriction fragment length polymorphism analysis and spoligotyping

被引:112
作者
Allix, Caroline
Walravens, Karl
Saegerman, Claude
Godfroid, Jacques
Supply, Philip
Fauville-Dufaux, Maryse
机构
[1] Inst Pasteur, Lab TB & Mycobacteries, B-1180 Brussels, Belgium
[2] Ctr Etud & Rech Vet & Agrochim, CODA, Sect Malad Bacteriennes & Immunol, B-1180 Brussels, Belgium
[3] Agence Fed Secur Chaine Alimentaire, DG Polit Controle, Secretariat Com Sci, B-1000 Brussels, Belgium
[4] Inst Pasteur, Lab Mecanismes Mol Pathogenese Bacterienne, INSERM, U629, F-59019 Lille, France
关键词
D O I
10.1128/JCM.01775-05
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Sources of Mycobacterium bovis contamination remain unclear for many cases of animal and human disease. A major limitation is the lack of sufficiently informative or epidemiologically well evaluated molecular methods for typing. Here, we report an evaluation of a high-throughput method based on 29 mycobacterial interspersed repetitive unit-variable-number tandem-repeat (MIRU-VNTR) loci to genotype 127 M. bovis isolates from cattle from 77 different Belgian farms, representative of a nationwide collection obtained from 1995 to 2003. MIRU-VNTR stability was demonstrated by analyzing a series of 74 isolates in total, obtained from different animals from a single farm or from different farms with an identified epidemiological link. The genotyping results and the genotypic diversity (h) were compared with those obtained by IS6110 restriction fragment length polymorphism (RFLP) analysis and spoligotyping. Among 68 isolates with no known epidemiological link, MIRU-VNTR typing discriminated better than either RFLP analysis or spoligotyping, with isolates taken individually (32 versus 16 and 17 genotypes; h = 0.91 versus 0.73 and 0.85, respectively) or in combination (32 versus 28 genotypes; h = 0.91 versus 0.92). Maximal resolution was already achieved with a subset of 9 loci. The observed congruence of the genetic relationships based on IS6110 RFLP analysis, spoligotyping, and MIRU-VNTR markers is consistent with a clonal population structure of M. bovis. These results support MIRU-VNTR typing as a convenient and discriminatory technique for analysis of the population structure of M. bovis in much greater detail and for addressing some still unresolved issues in the epidemiology of the pathogen.
引用
收藏
页码:1951 / 1962
页数:12
相关论文
共 34 条
[21]  
PALMER DNM, 2004, OIE MANUAL DIAGNOSTI
[22]   Investigation of the transmission of Mycobacterium bovis from deer to cattle through indirect contact [J].
Palmer, MV ;
Waters, WR ;
Whipple, DL .
AMERICAN JOURNAL OF VETERINARY RESEARCH, 2004, 65 (11) :1483-1489
[23]  
RIGOUTS LDM, 2000, 3 INT C MYC BOV, P39
[24]  
Robert J, 1999, INT J TUBERC LUNG D, V3, P711
[25]   Evaluation of variable number tandem repeat (VNTR) loci in molecular typing of Mycobacterium bovis isolates from Ireland [J].
Roring, S ;
Scott, AN ;
Hewinson, RG ;
Neill, SD ;
Skuce, RA .
VETERINARY MICROBIOLOGY, 2004, 101 (01) :65-73
[26]   Development of variable-number tandem repeat typing of Mycobacterium bovis:: Comparison of results with those obtained by using existing exact tandem repeats and spoligotyping [J].
Roring, S ;
Scott, A ;
Brittain, D ;
Walker, I ;
Hewinson, G ;
Neill, S ;
Skuce, R .
JOURNAL OF CLINICAL MICROBIOLOGY, 2002, 40 (06) :2126-2133
[27]   Discrimination of Mycobacterium tuberculosis complex bacteria using novel VNTR-PCR targets [J].
Skuce, RA ;
McCorry, TP ;
McCarroll, JF ;
Roring, SMM ;
Scott, AN ;
Brittain, D ;
Hughes, SL ;
Hewinson, RG ;
Neill, SD .
MICROBIOLOGY-SGM, 2002, 148 :519-528
[28]   The population structure of Mycobacterium bovis in Great Britain:: Clonal expansion [J].
Smith, NH ;
Dale, J ;
Inwald, J ;
Palmer, S ;
Gordon, SV ;
Hewinson, RG ;
Smith, JM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (25) :15271-15275
[29]   Identification of possible loci of variable number of tandem repeats in Mycobacterium tuberculosis [J].
Smittipat, N ;
Palittapongarnpim, P .
TUBERCLE AND LUNG DISEASE, 2000, 80 (02) :69-74
[30]   Variable human minisatellite-like regions in the Mycobacterium tuberculosis genome [J].
Supply, P ;
Mazars, E ;
Lesjean, S ;
Vincent, V ;
Gicquel, B ;
Locht, C .
MOLECULAR MICROBIOLOGY, 2000, 36 (03) :762-771