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Rapid, amplification-based fingerprinting of Mycobacterium tuberculosis
B B Plikaytis1, J T Crawford, C L Woodley
1Division of Bacterial and Mycotic Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30333.
Journal of General Microbiology
|July 1, 1993
Summary
A new gene amplification method, IS6110-ampliprinting, accurately types Mycobacterium tuberculosis strains by analyzing insertion element IS6110 locations. This method provides reliable patterns from sputum or cultures, correlating with IS6110 restriction fragment length polymorphism (RFLP) typing.
Area of Science:
- Microbiology
- Genetics
- Molecular Biology
Background:
- The insertion element IS6110 is present in multiple copies within the Mycobacterium tuberculosis genome.
- Variability in IS6110 insertion sites forms the basis for the IS6110 restriction fragment length polymorphism (RFLP) typing method.
Purpose of the Study:
- To introduce and evaluate a novel gene amplification method for assessing IS6110 location variability.
- To compare the discriminatory power of the new method with the established IS6110-RFLP typing.
Main Methods:
- A unilateral-nested polymerase chain reaction and hybridization procedure was employed.
- The method measured variable distances between IS6110 elements and a major polymorphic tandem repeat sequence in M. tuberculosis.
- The novel method was designated IS6110-ampliprinting.
Main Results:
- The generated amplicon patterns successfully clustered epidemiologically related M. tuberculosis strains.
- These clusters correlated well with groupings obtained through IS6110-RFLP typing.
- Reliable typing patterns could be generated directly from sputum specimens and M. tuberculosis cultures.
Conclusions:
- IS6110-ampliprinting is a reliable and effective gene amplification method for Mycobacterium tuberculosis strain typing.
- The method offers an alternative to IS6110-RFLP, with the advantage of direct application to clinical specimens.
- This technique aids in understanding the genomic variability of M. tuberculosis and its epidemiological implications.