Methodological issues in the quantification of subgingival microorganisms using the checkerboard technique.

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Dahlén, Gunnar,1944Gothenburg University,Göteborgs universitet,Institutionen för odontologi,Institute of Odontology (author)

Methodological issues in the quantification of subgingival microorganisms using the checkerboard technique.

Article/chapterEnglish2015

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Elsevier BV,2015

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LIBRIS-ID:oai:gup.ub.gu.se/215261
https://gup.ub.gu.se/publication/215261URI
https://doi.org/10.1016/j.mimet.2015.01.010DOI

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Language:English

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Subject category:art swepub-publicationtype

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The reproducibility and reliability of quantitative microbiological assessments using the DNA-DNA hybridization "checkerboard method" (CKB) were assessed. The data originated from 180 chronic periodontitis patients, who were enrolled in a clinical trial and sampled at baseline, and 3 and 12m post-therapy. The samples were divided into two portions allowing evaluation of reproducibility. In total, 531 samples were analyzed in a first run, using standard bacterial preparations of cells and 513 samples were accessible for analysis in the second, using standards based on purified DNA from the species. The microbial probe panel consisted of periodontitis marker bacteria as well as non-oral microorganisms. Three different ways of quantifying and presenting data; the visual scoring method, VSM, the standard curve method, SCM, and the percent method, PM, were compared. The second set of analyses based on the use of standard preparations of pure DNA was shown to be more consistent than the first set using standards based on cells, while the effect of storage time per se up to 2.5y seemed to be marginal. The best reproducibility was found for Tannerella forsythia, irrespective of quantification technique (Spearman's rho=0.587, Pearson's r≥0.540). The percent method (PM) based on percent of High Standard (10(6) cells) was more reliable than SCM based on a linear calibration of the High Standard and a Low Standard (10(5) cells). It was concluded that the reproducibility of the CBK method varied between different bacteria. High quality and pure specific DNA whole genomic probes and standards may have a stronger impact on the precision of the data than storage time and conditions.

Subject headings and genre

MEDICIN OCH HÄLSOVETENSKAP Klinisk medicin Odontologi hsv//swe
MEDICAL AND HEALTH SCIENCES Clinical Medicine Dentistry hsv//eng
MEDICIN OCH HÄLSOVETENSKAP Klinisk medicin hsv//swe
MEDICAL AND HEALTH SCIENCES Clinical Medicine hsv//eng

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Preus, H R (author)
Baelum, V (author)
Göteborgs universitetInstitutionen för odontologi (creator_code:org_t)

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In:Journal of microbiological methods: Elsevier BV110, s. 68-771872-83590167-7012

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MEDICAL AND HEALTH SCIENCES: MEDICAL AND HEAL ...; and Clinical Medicin ...; and Dentistry

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