Arbitrarily primed polymerase chain reaction as a rapid method to differentiate crossed from independent Pseudomonas cepacia infections in cystic fibrosis patients

Bingen, Édouard; Weber, Maximilian; Derelle, J.; Brahimi, Nima; Lambert-Zechovsky, N; Vidailhet, M.; Navarro, J; Élion, Jacques

doi:10.1128/jcm.31.10.2589-2593.1993

Cited by 51 publications

(25 citation statements)

References 31 publications

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“…PFGE provides a sensitive alternative approach for bacterial strain differentiation, molecular epidemiology and clonal analysis. Investigations by Bingen et al (3) have indicated that PFGE offered increased discriminatory power compared with AP-PCR and ribotyping in epidemiological studies of Pseudomonas cepacia. PFGE of Sacll digests of i^.…”

Section: Discussionmentioning

confidence: 99%

“…Arbitrarily primed polymerase chain reaction (AP-PCR) has proven to be a valuable tool in studies of genetic polymorphisms, epidemiological investigations, clonal analysis and studies of the population genetics of a variety of organisms (3,14,17,19,20,25). AP-PCR was originally described by Welsh & McClelland (30) and Williams et al (31) as a method for analyzing genetic polymorphisms of both prokaryotic and eukaryotic cells.…”

mentioning

confidence: 99%

“…The greatest advantage of this technique is the ability to provide specific DNA fingerprints without prior knowledge of the DNA sequence of the organism (30,31). Pulsed field gel electrophoresis (PFGE) has also proven to be a valuable tool in bacterial strain differentiation and clonal analysis (1,3,21,27). PFGE is a variation of restriction endonuclease analysis that examines DNA fragments larger than 10 kb.…”

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confidence: 99%

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Arbitrarily primed polymerase chain reaction fingerprinting and clonal analysis of oral Fusobacterium nucleatum isolates

George

Reynolds

Falkier

1997

Oral Microbiology and Immunology

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F. nucleatum is the most commonly isolated microorganism from subgingival plaque, but the role of this microorganism in periodontal diseases remains undefined. Arbitrarily primed polymerase chain reaction (AP-PCR) was evaluated as a method for fingerprinting F. nucleatum isolates and for use in clonal analysis. Pulsed field gel electrophoresis was used to further differentiate F. nucleatum isolates, with identical AP-PCR patterns. Extremely heterogeneous AP-PCR fingerprints were observed among the 98 F. nucleatum isolates, with 36 different genotypes observed with primer C1 and 30 different genotype detected with primer C2. Combining the results of the AP-PCR genotype analysis from C1 and C2 primer amplifications revealed that up to 7 different genotypes could be distinguished from isolates from the same oral cavity and that up to 4 different genotypes were observed within a single site. An intense amplicon at approximately 450 bp generated in AP-PCR amplification with primer C2 was associated with F. nucleatum subsp. nucleatum (ATCC 25586) and with 15 F. nucleatum isolates from diseased sites and 2 isolates from healthy sites. Pulsed field gel electrophoresis confirmed the AP-PCR genotypes and demonstrated increased discriminatory power over AP-PCR. The results indicated that AP-PCR and pulsed field gel electrophoresis provide a simple and sensitive means for differentiating oral F. nucleatum isolates and further demonstrate the heterogeneity of this species. These techniques may serve as useful tools in the clonal and epidemiological analysis of F. nucleatum isolates, which may help define the role of these microorganisms in periodontal diseases.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Arbitrarily primed polymerase chain reaction fingerprinting and clonal analysis of oral Fusobacterium nucleatum isolates

George

Reynolds

Falkier

1997

Oral Microbiology and Immunology

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“…By summing the results of AP-PCR profiles generated with three different primers, Saulnier et al (10) were able to identify 25 AP-PCR patterns in a group of 26 strains of Staphylococcus aureus shown to be different by CHEF. Bingen et al (4) used AP-PCR to identify four different strains of Pseudomonas cepacia in a group of 23 isolates, whereas CHEF was able to discriminate five strains. Similarly, in our study, the improvement in sensitivity of CHEF over AP-PCR was a small one, identifying 49 CHEF patterns in a group of isolates having 47 AP-PCR patterns.…”

Section: Discussionmentioning

confidence: 99%

Genomic fingerprinting of epidemic and endemic strains of Stenotrophomonas maltophilia (formerly Xanthomonas maltophilia) by arbitrarily primed PCR

et al. 1995

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Arbitrarily primed PCR (AP-PCR) was used to type 64 clinical isolates of Stenotrophomonas maltophilia from 60 patients and the hands of one nurse. Forty-seven different patterns were observed, most patients having isolates with unique genomic fingerprints. A single pattern, however, was obtained from six of eight patients involved in an intensive care nursery outbreak, confirming the suspected nosocomial transmission of this microorganism. This strain was also found in four other patients hospitalized at the same time but in different units. AP-PCR typing results had a good correlation with the 49 patterns obtained when the isolates were typed by contour-clamped homogeneous electric field gel electrophoresis. Although AP-PCR is slightly less discriminatory than contour-clamped homogeneous electric field gel electrophoresis, it offers several advantages and should be considered as a practical option for molecular typing during investigations of outbreaks.

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“…ERIC sequences are often used as primers, both for Gram-positive bacteria such as S. aureus (32), and for Gram-negative species such as Stenotrophomonas maltophilia (33). For B. cepacia, promising results indicate equal resolution power with AP-PCR and ribotyping, and only minimally less discriminative power compared with PFGE (34).…”

Section: Discussionmentioning

confidence: 99%

Arbitrarily primed PCR and sequencing of 16S rDNA for epidemiological typing and species identification of Burkholderia cepacia isolates from Swedish patients with cystic fibrosis reveal genetic heterogeneity^Note

Karpati

Giedraitis

Thore

et al. 2001

APMIS

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To investigate whether arbitrarily primed (AP)-PCR and/or 16S rDNA sequencing could be used as rapid methods for epidemiological typing and species identification of clinical Burkholderia isolates from patients with cystic fibrosis (CF), a total of 39 clinical B. cepacia isolates, including 33 isolates from 14 CF patients, were fingerprinted. ERIC-2 primer was used for AP-PCR. The AP-PCR clustering analysis resulted in 14 different clusters at a 70% similarity level. The AP-PRC patterns were individual despite considerable similarities. To sequence rDNA, a broad-range PCR was applied. The PCR product included four variable loops (V8, V3, V4 and V9) of the 16S ribosomal small subunit RNA gene. The multiple sequence alignment produced 12 different patterns, 5 of them including more than one isolate. Heterogeneity of the bases in the V3 region, indicating the simultaneous presence of at least two different types of 16S rRNA genes in the same cell, was revealed in 10 isolates. Most of the CF patients were adults who had advanced disease at follow-up. Both the sequencing and the AP-PCR patterns revealed genetic heterogeneity of isolates between patients. According to the results obtained, AP-PCR could advantageously be used for epidemiological typing of Burkholderia, whereas partial species identification could effectively be obtained by sequencing of the V3 region of the 16S RNA gene.

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Arbitrarily primed polymerase chain reaction as a rapid method to differentiate crossed from independent Pseudomonas cepacia infections in cystic fibrosis patients

Cited by 51 publications

References 31 publications

Arbitrarily primed polymerase chain reaction fingerprinting and clonal analysis of oral Fusobacterium nucleatum isolates

Arbitrarily primed polymerase chain reaction fingerprinting and clonal analysis of oral Fusobacterium nucleatum isolates

Genomic fingerprinting of epidemic and endemic strains of Stenotrophomonas maltophilia (formerly Xanthomonas maltophilia) by arbitrarily primed PCR

Arbitrarily primed PCR and sequencing of 16S rDNA for epidemiological typing and species identification of Burkholderia cepacia isolates from Swedish patients with cystic fibrosis reveal genetic heterogeneity^Note

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Arbitrarily primed polymerase chain reaction as a rapid method to differentiate crossed from independent Pseudomonas cepacia infections in cystic fibrosis patients

Cited by 51 publications

References 31 publications

Arbitrarily primed polymerase chain reaction fingerprinting and clonal analysis of oral Fusobacterium nucleatum isolates

Arbitrarily primed polymerase chain reaction fingerprinting and clonal analysis of oral Fusobacterium nucleatum isolates

Genomic fingerprinting of epidemic and endemic strains of Stenotrophomonas maltophilia (formerly Xanthomonas maltophilia) by arbitrarily primed PCR

Arbitrarily primed PCR and sequencing of 16S rDNA for epidemiological typing and species identification of Burkholderia cepacia isolates from Swedish patients with cystic fibrosis reveal genetic heterogeneityNote

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Arbitrarily primed PCR and sequencing of 16S rDNA for epidemiological typing and species identification of Burkholderia cepacia isolates from Swedish patients with cystic fibrosis reveal genetic heterogeneity^Note