Evaluation of alternatives to RFLP for the analysis of clustered cases of tuberculosis

Int J Tuberc Lung Dis. 2006 Apr;10(4):454-9.

Abstract

Background: Mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) analysis is a recently developed method which could be suitable as a 'real-time' genotyping tool for Mycobacterium tuberculosis.

Methods: One hundred and thirty-four M. tuberculosis isolates were analysed using the reference method, IS6110-restriction fragment length polymorphism (RFLP), and by MIRU, alone and together with spoligotyping.

Results: MIRU reduced the genotyping turnaround time by 21 days. The discriminatory power (HGDI) for MIRU and RFLP was 0.978 and 0.989, respectively. RFLP clustered 41.8% of the isolates (17 clusters; 2-9 representatives), whereas MIRU increased the number and size of the clusters (57.5% of the isolates in 20 clusters; 2-14 representatives). With respect to the RFLP clusters, MIRU data showed full correlation in only 7/ 17 (41%) clusters and low correlation in 8/17 (47%) clusters. When MIRU and spoligotyping were considered together, the analysis fitted better with RFLP data: 1) 42.5% of the isolates were grouped in 20 clusters of 2-6 representatives, and 2) the number of clusters with full correlation with RFLP data increased to 11/17 and those with low correlation decreased to 2/17.

Conclusion: MIRU-VNTR analysis showed low correlation with RFLP. The addition of spoligotyping to MIRU analysis fitted much better with RFLP analysis, although full correlation was still not achieved.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Bacterial / analysis*
  • Genotype
  • Humans
  • In Vitro Techniques
  • Mycobacterium tuberculosis / genetics*
  • Mycobacterium tuberculosis / isolation & purification
  • Polymorphism, Restriction Fragment Length*
  • Reproducibility of Results
  • Tuberculosis / diagnosis
  • Tuberculosis / microbiology*

Substances

  • DNA, Bacterial