Genomic fingerprinting of "Haemophilus somnus" isolates by using a random-amplified polymorphic DNA assay

Myers, Leslie; Silva, S. V. P. S.; Procunier, J. D.; Little, P B

doi:10.1128/jcm.31.3.512-517.1993

Cited by 39 publications

(18 citation statements)

References 25 publications

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“…AP-PCR has been used for fingerprinting individual strains of various microbial species (1,17,21,22). Also, certain AP-PCR banding patterns have been able to distinguish between microbial species (19). The present study demonstrated the usefulness of AP-PCR to distinguish individual B. forsythus strains.…”

Section: Discussionsupporting

confidence: 51%

Molecular detection of Bacteroides forsythus in human periodontitis

Lotufo

Flynn

Chen

et al. 1994

Oral Microbiology and Immunology

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The usefulness of a digoxigenin-labeled genomic DNA probe for the detection of subgingival Bacteroides forsythus was examined. In addition, the arbitrarily primed polymerase chain reaction (AP-PCR) was used to delineate the genetic diversity of B. forsythus periodontal isolates. The DNA probe detected 10(3) B. forsythus cells and yielded a strong signal at 10(4) cells. It reacted with B. forsythus ATCC 43037T and 44 clinical isolates and showed no detectable reactivity with 75 strains of 24 other oral microbial species. In comparison to culture, the DNA probe in a dot-blot method demonstrated a sensitivity of 88.8% and a specificity of 38.4% (accuracy, 72.5%). By colony-blotting on primary plates, a sensitivity of 98.1% and a specificity of 53.8% (accuracy, 82.5%) were obtained. B. forsythus was detected in 449 (73.1%) of 614 periodontitis patients. The occurrence of the organism was closely associated with Porphyromonas gingivalis, both species being present in 54.8% and absent in 22.2% of 270 study samples. AP-PCR identified 24 B. forsythus genotypes among 27 test strains. This study demonstrated the utility of a non-radioactive genomic probe for direct detection of B. forsythus in subgingival specimens. The species showed a considerable degree of genetic diversity. DNA analysis may help to determine the role of B. forsythus in periodontal disease and its mode of transmission among exposed individuals.

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Section: Discussionsupporting

confidence: 51%

Molecular detection of Bacteroides forsythus in human periodontitis

Lotufo

Flynn

Chen

et al. 1994

Oral Microbiology and Immunology

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“…However, since the PCR tnethod is sensitive to variation in tetnperature profiles and to changes in concentrations of genotnic Size (kb) ;' 23,1 -9.4 -6.5 • 4.3 . (12,21) and strains of tnedically itnportant bacterial species (3,11,14). AP-PCR was able to distinguish H. .sotnttus frotn other bacterial species and to identify individual H. .sointius strains (14), Several virulent H. sonmus strains were found to possess tiearly identical AP-PCR DNA fingerprints, suggesting a clonal origin of virulent straitis within this species, Iti additioti, the genetic relatedness between //.…”

Section: Discussionmentioning

confidence: 94%

“…(12,21) and strains of tnedically itnportant bacterial species (3,11,14). AP-PCR was able to distinguish H. .sotnttus frotn other bacterial species and to identify individual H. .sointius strains (14), Several virulent H. sonmus strains were found to possess tiearly identical AP-PCR DNA fingerprints, suggesting a clonal origin of virulent straitis within this species, Iti additioti, the genetic relatedness between //. sotnnus strains could be detertnined frotn the sitnilarity coefficient calculated frotn pairwise cotnparison of the DNA fingerprints generated by AP-PCR (14).…”

Section: Discussionmentioning

confidence: 99%

“…The resulting amplification products, consisting of various DNA fragtnents, tnay serve as polymorphic markers. AP-PCR has been used for getietic analysis of the genus Candida (11), for DNA fingerprinting of Haetnophilus .sotntms (14), for generating polytnorphistn tnarkers linked to specific genes or genomic regions in plants (13), for the study of population genetics (6) and for gene mapping (15,20). The aitn of the present study was to determine the utility of AP-PCR for clonal analysis of A. aetitwmyeetetneotnitans clinical isolates and to compare the results with those of an established Southern blot analysis using a cloned A. actinotnyeetetneotnitans DNA fragment.…”

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confidence: 99%

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Evaluating two methods for fingerprinting genomes of Actinobacillus actinomycetemcomitans

Slots

Liu

DiRienzo

et al. 1993

Oral Microbiology and Immunology

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The arbitrary primer polymerase chain reaction (AP-PCR) and Southern blot restriction fragment length polymorphism (RFLP) were used to genotype the periodontal pathogen A. actinomycetemcomitans. Total genomic DNA from 73 strains was extracted by conventional methods. Three random-sequence 10-base oligonucleotide primers were chosen for AP-PCR. The amplified DNA products were separated electrophoretically in a 1% agarose gel containing ethidium bromide and the banding patterns were compared among different strains. For RFLP analysis, DNA was digested with EcoRI, separated on a 0.8% agarose gel and transferred to a nylon membrane. The membrane was probed with a previously characterized 5.2 kilobases (kb) DNA fragment cloned from A. actinomycetemcomitans strain Y4. The probe was labeled with digoxigenin, and hybridized fragments were detected with anti-digoxigenin antibody. AP-PCR produced 4-10 DNA bands in the 0.5-5 kb regions and distinguished 9, 13 or 17 genotypes, depending on the specific primer used. Southern blot RFLP analysis revealed 12 hybridization patterns consisting of 1 or 2 DNA fragments (2-23 kb). The addition of the Southern blot analysis to the AP-PCR analysis gave rise to a total of 30 DNA profiles among the 73 A. actinomycetemcomitans study strains. The results indicate that both AP-PCR and Southern blot analysis are useful in clonal analysis of A. actinomycetemcomitans.

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“…28 AP-PCR has been used for fingerprinting individual strains of various microbial species, [29][30][31] and banding patterns have been used to distinguish between microbial species. 32 In our study, the AP-PCR method showed a great polymorphism among F. nucleatum species. However, a poor degree of technical reproducibility and discriminatory power was achieved by this typing method in accordance with George et al 11 Several parameters can alter AP-PCR profiles and make standardization very difficult to obtain reproducible results.…”

Section: Discussionmentioning

confidence: 48%

Arbitrarily Primed‐Polymerase Chain Reaction for Identification and Epidemiologic Subtyping of Oral Isolates of Fusobacterium nucleatum

Ávila-Campos

Sacchi

Whitney

et al. 1999

Journal of Periodontology

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Our results suggest that PCR primer pairs 5059S, FN5047 or M1211 can be used to specifically identify F. nucleatum isolates and distinguish them from other bacteria. The primer pair M8171 could also be used to differentiate F. nucleatum isolated from periodontal patients or healthy individuals. These specific primers can be used in PCR analysis for specific identification of F. nucleatum and to distinguish it from other bacteria associated with human periodontitis. These approaches appear promising in facilitating laboratory identification, molecular subtyping, and taxonomy of putative periodontopathogens.

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Genomic fingerprinting of "Haemophilus somnus" isolates by using a random-amplified polymorphic DNA assay

Cited by 39 publications

References 25 publications

Molecular detection of Bacteroides forsythus in human periodontitis

Molecular detection of Bacteroides forsythus in human periodontitis

Evaluating two methods for fingerprinting genomes of Actinobacillus actinomycetemcomitans

Arbitrarily Primed‐Polymerase Chain Reaction for Identification and Epidemiologic Subtyping of Oral Isolates of Fusobacterium nucleatum

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