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Related Experiment Videos

Molecular methods for Mycobacterium tuberculosis strain typing: a users guide.

E Kanduma1, T D McHugh, S H Gillespie

  • 1Clinical Laboratory, Kilimanjaro Christian Medical College, PO Box 3010, Moshi, Tanzania.

Journal of Applied Microbiology
|April 16, 2003
PubMed
Summary
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Choosing the right Mycobacterium tuberculosis typing technique is crucial. IS6110 restriction fragment length polymorphism (RFLP) is the gold standard, but mycobacterial interspersed repetitive unit (MIRU) typing shows promise for future epidemiological studies.

Area of Science:

  • Microbiology
  • Epidemiology
  • Molecular Biology

Background:

  • Mycobacterium tuberculosis (M.tb.) typing is essential for disease control.
  • Various molecular techniques exist, but selecting the optimal method presents challenges.

Purpose of the Study:

  • To review and compare available M.tb. typing techniques.
  • To guide the selection of appropriate methods based on study scale and objectives.

Main Methods:

  • IS6110 restriction fragment length polymorphism (RFLP) as the current gold standard for large-scale studies.
  • Mycobacterial interspersed repetitive unit (MIRU) typing, including MIRU-variable number of tandem repeats (VNTR), as an emerging standard.
  • Polymorphic Guanine cytosine-rich repetitive sequence (PGRS) for low copy number strains and cluster confirmation.

Related Experiment Videos

  • PCR-based methods for rapid initial investigation of local outbreaks and contamination.

    • Main Results:

    • IS6110 RFLP offers portability and standardization for epidemiological studies.
    • MIRU-VNTR provides superior reproducibility and discrimination, favored for its performance.
    • PGRS is effective for typing low copy number strains and confirming true clusters.
    • PCR-based methods are valuable for speed in outbreak investigations.

    Conclusions:

    • The choice of M.tb. typing technique depends on the specific application, from large-scale epidemiology to rapid outbreak detection.
    • MIRU-VNTR is poised to become a key method, potentially challenging IS6110 RFLP.
    • Confirmatory typing using IS6110 RFLP or PGRS is recommended when MIRU-VNTR matches are found.