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Related Concept Videos

Modern Molecular Taxonomy01:29

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Advancements in molecular biology have revolutionized the identification and characterization of bacteria, with multiple methods leveraging DNA sequencing for enhanced precision. As sequencing technologies improve and costs decline, these approaches are increasingly used in clinical, environmental, and evolutionary studies.Multilocus Sequence Typing (MLST) examines several housekeeping genes, essential chromosomal genes encoding cellular functions, to distinguish strains. Approximately...
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Demonstrating a Multi-drug Resistant Mycobacterium tuberculosis Amplification Microarray
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Published on: April 25, 2014

Typing Mycobacterium tuberculosis using variable number tandem repeat analysis.

T J Brown1, V N Nikolayevskyy, F A Drobniewski

  • 1HPA MRU, Queen Mary's School of Medicine and Dentistry, 2 Newark Street, London E1 2AT, UK. t.brown@qmul.ac.uk

Methods in Molecular Biology (Clifton, N.J.)
|June 19, 2010
PubMed
Summary
This summary is machine-generated.

Variable number tandem repeat (VNTR) analysis offers a powerful DNA-based method for studying Mycobacterium tuberculosis evolution and epidemiology. This technique provides high discrimination for both evolutionary and epidemiological investigations.

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Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis

Published on: June 17, 2019

Area of Science:

  • Microbiology
  • Genetics
  • Epidemiology

Background:

  • DNA-based typing is crucial for understanding Mycobacterium tuberculosis (M. tuberculosis) epidemiology and evolution.
  • IS6110 RFLP, an early method, faced technological challenges and had limited use in evolutionary studies.
  • PCR-based methods like spoligotyping and VNTR analysis offer improved solutions.

Purpose of the Study:

  • To describe methods for Variable Number Tandem Repeat (VNTR) analysis for M. tuberculosis typing.
  • To evaluate the utility of VNTR analysis in both epidemiological and evolutionary studies.
  • To present both high-throughput automated and manual agarose gel-based methods for VNTR analysis.

Main Methods:

  • Amplification of PCR fragments for multiple VNTR loci.
  • Determination of PCR fragment sizes to calculate the number of tandem repeats.
  • Utilizing a numerical code based on repeat numbers at various loci to define a VNTR type.
  • Description of a high-throughput automated method using capillary fragment analysis for 15 loci.
  • Description of a manual method employing agarose gel electrophoresis.

Main Results:

  • VNTR analysis, particularly using panels of selected loci, demonstrates utility in both epidemiological and evolutionary studies of M. tuberculosis.
  • Spoligotyping alone shows limited discriminatory power for epidemiology but is useful for evolutionary insights.
  • VNTR typing provides a numerical code for classifying bacterial types based on repeat numbers.

Conclusions:

  • VNTR analysis is a valuable tool for M. tuberculosis research, offering high discrimination for epidemiological and evolutionary investigations.
  • Both automated and manual VNTR methods are presented, catering to different laboratory capabilities.
  • VNTR typing significantly advances the ability to track and understand M. tuberculosis strains.