Numerical Analysis of Normalized Whole-cell Protein Profiles after Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis

Plikaytis, Brian D.; Carlone, George M.; Plikaytis, Bonnie B.

doi:10.1099/00221287-132-9-2653

Cited by 16 publications

(21 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, a window of 2.6 mm was used for band matching based on 2xSD of the most variable band evaluated. Similarities between strains were based on matching co-migrating band positions, between pairs of band profiles using the Dice coefficient as described by Plikaytis et al (18). Two bands were considered to be a match if they were within a window of 2.6 mm of each other.…”

Section: Discussionmentioning

confidence: 99%

Detection of identical ribotypes of Porphyromonas gingivalis in patients residing in the United States, Sudan, Romania and Norway

Ali¹,

Martin²,

Socransky³

1997

Oral Microbiology and Immunology

View full text Add to dashboard Cite

Porphyromonas gingivalis has been isolated from periodontitis lesions in subjects from many geographical locations. The purpose of this investigation was to determine whether similar ribotypes of P. gingivalis could be detected among strains isolated in different countries. A total of 198 isolates of P. gingivalis were obtained from 52 periodontitis patients in Boston (130 isolates), Bergen, Norway (17 isolates), Khartoum, Sudan (26 isolates), and Bucharest, Romania (25 isolates). DNA was isolated from each strain, cut separately by the restriction endonucleases KpnI and PstI. The resulting preparations were subjected to electrophoresis in a 0.8% agarose gel using a Tris-acetate EDTA buffer. Uncut lambda and a 1000-bp fragment of 16S rRNA were included as internal standards in each lane. In addition, a HindIII digest of lambda was present in a separate lane in each run. The DNA fragments were transferred to a nylon membrane by downward capillary transfer. 16S rRNA bands were detected using a 1000-kb digoxigenin-labelled probe generated by a polymerase chain reaction. At the same time, a digoxigenin-labelled probe to lambda was employed to detect the internal and molecular weight standards. The bands were detected using antibody to digoxigenin conjugated to alkaline phosphatase and chemiluminescence. The positions of the bands relative to the internal standards were determined and normalized to correct for run-to-run variations, and the molecular weight of each band was determined by comparison with standards within each gel. The resulting data for the 2 enzymes were combined and subjected to cluster analysis using an average unweighted linkage sort. In some instances, isolates that appeared to be of identical ribotype using one endonuclease gave different ribotypes using the other. Strains of P. gingivalis within a subject were usually identical, except for 3 patients who harbored 2 different ribotypes/individual. All subsequent analyses employed a single ribotype strain for each subject. A total of 32 ribotypes were observed for isolates from distant countries. A total of 11.5% of the patients had isolates exhibiting the same ribotype: ribotype 7a. Identical ribotypes of P. gingivalis can be recovered from subgingival plaque samples of periodontitis patients in different countries.

show abstract

Section: Discussionmentioning

confidence: 99%

Detection of identical ribotypes of Porphyromonas gingivalis in patients residing in the United States, Sudan, Romania and Norway

Ali¹,

Martin²,

Socransky³

1997

Oral Microbiology and Immunology

View full text Add to dashboard Cite

show abstract

“…To overcome this, other phenotypic and genotypic techniques have been developed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) has gained ground during the latest years in differentiating and classifying microorganisms (Plikaytis et al 1986). It is a sensitive and relatively simple method applicable to a wide range of microorganisms (Coenye et al 2001).…”

Section: Introductionmentioning

confidence: 99%

Comparative typing of Pseudomonas species isolated from the aquatic environment in Greece by SDS-PAGE and RAPD analysis

et al. 2005

View full text Add to dashboard Cite

E . S A Z A K L I , M . L E O T S I N I D I S , A . V A N T A R A K I S A N D M . P A P A P E T R O P O U L O U . 2005.Aims: Three broadly used typing methods were employed in order to assess and compare the identification and classification of environmental Pseudomonas strains. The reproducibility, typeability and discriminatory power of the methods were also compared to evaluate their application. Finally, the potential impact on public health of the isolates is to be discussed. Methods and Results: Pseudomonas strains (160) isolated from the aquatic environment in Greece and identified by a rapid identification commercially available system (API20NE), were subjected to whole-cell protein electrophoresis (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and Randomly Amplified Polymorphic DNAs (RAPD) using two 10-mer primers. In general, the obtained results were in agreement. Twenty isolates that could not be identified by the API20NE system were classified by the other methods. Conclusions: Rapid identification systems may serve only for a first rough identification of environmental Pseudomonads. In order to acquire further information, so that conclusions about their role in the ecosystem and human health could be drawn, other phenotypic or genotypic methods have to be applied. Significance and Impact of Study: It is important, from a public health point of view, to monitor the identities of environmental Pseudomonas isolates using specific methods due to their ubiquity, heterogeneity and their pathogenicity, either established or potential.

show abstract

“…In the present study, migration distances, determined by digitizing gels, were converted to fragment sizes based on a 1-kb ladder standard before computerized analysis for genetic relatedness. Although there have been several attempts to use densitometry, or soft-laser scanning, to convert restriction digest patterns or protein profiles to quantitative data on the basis of inclusion of internal or external standards (2,5,18,21,25,32), these methods were not practical for use in our experiments since our application of ribotyping to many species of bacteria resulted in wide ranges of fragment distribution. Previously described standards would potentially conflict with patterns obtained.…”

Section: Discussionmentioning

confidence: 99%

Restriction fragment length polymorphisms in the ribosomal genes for species identification and subtyping of aerotolerant Campylobacter species

et al. 1991

View full text Add to dashboard Cite

Whole-cell chromosomal digests of 84 strains of aerotolerant Campylobacter (AC) were examined by using PvuII restriction fragment length polymorphisms of rRNA genes followed by hybridization with Escherichia coli 16S and 23S rRNA (ribotyping). The AC strains belonged to Campylobacter cryaerophila (n = 13) and a newly defined species, "C. butzleri" (n = 64). Strains of C. cryaerophila belonged to two hybridization groups: DNA group 1A (including the type strain of C. cryaerophila) and DNA group 1B (J. A

show abstract

Numerical Analysis of Normalized Whole-cell Protein Profiles after Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis

Cited by 16 publications

References 15 publications

Detection of identical ribotypes of Porphyromonas gingivalis in patients residing in the United States, Sudan, Romania and Norway

Detection of identical ribotypes of Porphyromonas gingivalis in patients residing in the United States, Sudan, Romania and Norway

Comparative typing of Pseudomonas species isolated from the aquatic environment in Greece by SDS-PAGE and RAPD analysis

Restriction fragment length polymorphisms in the ribosomal genes for species identification and subtyping of aerotolerant Campylobacter species

Contact Info

Product

Resources

About