Document Detail

Genetic diversity among strains of Moraxella catarrhalis: analysis using multiple DNA probes and a single-locus PCR-restriction fragment length polymorphism method.
MedLine Citation:
PMID:  9650948     Owner:  NLM     Status:  MEDLINE    
Moraxella (Branhamella) catarrhalis, a causative agent of otitis media, sinusitis, and exacerbation of bronchitis, has acquired widespread ability to produce beta-lactamase and can be nosocomially transmitted. The typing methods used in epidemiological analyses of M. catarrhalis are not optimal for genetic analyses. Two methods, a multiple-locus Southern blot (SB) method and a single-locus PCR-restriction fragment length polymorphism (RFLP) method, were developed and used to assess genetic diversity and potential clinical and geographic relationships in M. catarrhalis. Nine randomly cloned M. catarrhalis DNA fragments were used as probes of SBs containing DNA from 54 geographically and clinically diverse strains. For comparison, a PCR-RFLP method was developed as a quick, inexpensive, and discriminating alternative. A highly variable 3.7-kb genomic region (M46) was cloned and sequenced, and 3.5 kb of the cloned DNA was targeted for PCR amplification. DNAs from the 54 strains were subjected to PCR-RFLP. SB analysis distinguished all strains that had no apparent epidemiological linkage (40 of 54), and PCR-RFLP distinguished fewer strains (21 of 54). Epidemiologically linked strains appeared genetically identical by both methods. PCR-RFLP was compared to pulsed-field gel electrophoresis (PFGE) for 8 of the 54 strains and 23 additional strains. PCR-RFLP distinguished fewer strains than PFGE typing (16 of 31 versus 20 of 31 strains), but PCR-RFLP was more useful for inferring interstrain relatedness. Separate cluster analyses of multilocus SB and single locus PCR-RFLP data showed high genetic diversity within and across geographic locations and clinical presentations. The resultant dendrograms were not entirely concordant, but both methods often gave similar strain clusters at the terminal branches. High genetic diversity, nonconcordance of cluster analyses from different genetic loci, and shared genotypes among epidemiologically linked strains support a hypothesis of high recombination relative to spread of clones. Single-locus PCR-RFLP may be suitable for short-term epidemiological studies, but the SB data demonstrate that greater strain discrimination may be obtained by sampling variation at multiple genomic sites.
E S Walker; R A Preston; J C Post; G D Ehrlich; J H Kalbfleisch; K L Klingman
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of clinical microbiology     Volume:  36     ISSN:  0095-1137     ISO Abbreviation:  J. Clin. Microbiol.     Publication Date:  1998 Jul 
Date Detail:
Created Date:  1998-09-28     Completed Date:  1998-09-28     Revised Date:  2013-04-18    
Medline Journal Info:
Nlm Unique ID:  7505564     Medline TA:  J Clin Microbiol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1977-83     Citation Subset:  IM    
James H. Quillen Veterans Affairs Medical Center, Department of Internal Medicine, Quillen College of Medicine, East Tennessee State University, Johnson City, USA.
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MeSH Terms
Bacterial Typing Techniques
Blotting, Southern
Cloning, Molecular
Cluster Analysis
DNA Restriction Enzymes
DNA, Bacterial / analysis
Data Collection
Electrophoresis, Gel, Pulsed-Field
Genetic Variation*
Molecular Sequence Data
Moraxella (Branhamella) catarrhalis / classification,  genetics*,  isolation & purification
Neisseriaceae Infections / epidemiology
Polymerase Chain Reaction / methods*
Polymorphism, Restriction Fragment Length*
Sequence Analysis, DNA
Grant Support
Reg. No./Substance:
0/DNA, Bacterial; EC 3.1.21.-/DNA Restriction Enzymes

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