An IS 6110-targeting fluorescent amplified fragment length polymorphism alternative to IS 6110 restriction fragment length polymorphism analysis for Mycobacterium tuberculosis DNA fingerprinting

N. Thorne, J. T. Evans, E. G. Smith, P. M. Hawkey, Saheer Gharbia, Catherine Arnold*

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    8 Citations (Scopus)

    Abstract

    A rapid, simple and highly discriminatory DNA fingerprinting methodology which produces data that can be easily interpreted, compared and transported is the ultimate goal for studying the epidemiology of Mycobacterium tuberculosis . A novel Taq I fluorescent amplified fragment length polymorphism (fAFLP) approach to M. tuberculosis DNA fingerprinting that targeted the variable IS 6110 marker was developed in this study. The new method was tested for specificity and reproducibility, and compared with the standard reference IS 6110 restriction fragment length polymorphism (RFLP) method for a panel of 78 isolates. Clustering conflicts between the two methods were resolved using mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) data. Comparison with an in-silico digestion of strain H37Rv showed that fAFLP-detected fragments were highly specific in vitro . The reproducibility of repeated digestions of strain H37Rv was 100%. Clustering results obtained by fAFLP and RFLP were highly congruent, with fAFLP allocating 97% of RFLP-clustered isolates to the same eight clusters as RFLP. Two single-copy isolates that had been clustered by RFLP were not clustered by fAFLP, but the MIRU-VNTR patterns of these isolates were different, indicating that the RFLP data had falsely clustered these isolates. Analysis by fAFLP will allow rapid screening of isolates to confirm or refute epidemiological links, and thereby provide insights into the frequency, conservation and consequences of specific transposition events.

    Original languageEnglish
    Pages (from-to)964-970
    Number of pages7
    JournalClinical Microbiology and Infection
    Volume13
    Issue number10
    DOIs
    Publication statusPublished - Oct 2007

    Keywords

    • Amplified fragment length polymorphism (AFLP) analysis
    • Fluorescent AFLP analysis
    • IS6110
    • Molecular typing
    • Mycobacterium tuberculosis
    • RFLP
    • Typing

    Fingerprint

    Dive into the research topics of 'An IS 6110-targeting fluorescent amplified fragment length polymorphism alternative to IS 6110 restriction fragment length polymorphism analysis for Mycobacterium tuberculosis DNA fingerprinting'. Together they form a unique fingerprint.

    Cite this