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PMID: 8099087 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mixed-linker polymerase chain reaction: a new method for rapid fingerprinting of isolates of the Mycobacterium tuberculosis complex.

Journal of clinical microbiology ·Vol. 31 ·No. 5 ·1993-05-00 ·Pages 1293-8

Haas WH, Butler WR, Woodley CL, Crawford JT

Abstract

Rapid recognition of multidrug-resistant strains of Mycobacterium tuberculosis is a desirable goal for treatment of patients and protection of health care workers. DNA fingerprints produced with the insertion sequence IS6110 generate restriction fragment length polymorphism (RFLP) patterns that reliably identify M. tuberculosis complex strains. This report describes a rapid technique for RFLP typing using the polymerase chain reaction. The method uses one primer specific for IS6110 and a second primer complementary to a linker ligated to the restricted genomic DNA. In one strand the linker contains uracil in place of thymidine, and specific amplification is obtained by elimination of this strand with uracil N-glycosylase. Mixed-linker fingerprinting clearly differentiated multidrug-resistant isolates from 12 outbreaks and unambiguously assigned them to 26 RFLP groups.

MeSH Terms
Base Sequence DNA Fingerprinting/methods DNA Transposable Elements DNA, Bacterial/genetics Drug Resistance, Microbial Evaluation Studies as Topic Humans Molecular Sequence Data Mycobacterium tuberculosis/drug effects,genetics,isolation & purification Polymerase Chain Reaction/methods Polymorphism, Restriction Fragment Length Software Design
Chemicals
DNA Transposable Elements DNA, Bacterial
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Haas W H
National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333.
Butler W R
Woodley C L
Crawford J T
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Article Info
Journal
Journal of clinical microbiology
Abbr.
J Clin Microbiol
ISSN
0095-1137
DOI
10.1128/jcm.31.5.1293-1298.1993
Published
1993-05-00
Pages
1293-8
Language
English
Region
United States
NLM ID
7505564
PMCID
PMC262921
Subset
IM
External Links
PubMed source DOI: 10.1128/jcm.31.5.1293-1298.1993 PMC Full Text View journal details
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