A rapid solid-phase fluorimetric assay for measuring bacterial adherence, using DNA-binding stains.
In this report, we describe the validation of a rapid, single-step, microtiter plate method for quantifying bacterial adherence, based on fluorescent labeling of microorganisms with cell-permeable fluorescent DNA-binding probes. We have tested the binding to saliva-coated microtiter plates of bacteria, including Helicobacter pylori and viridans streptococci (S. mitis, S. gordonii, S. sanguis), known to interact with salivary components. Furthermore, we tested the short-term and longer-term temporal stability of a saliva-mediated adherence of these bacteria in a healthy population (N=30). The assay exhibited excellent reliability statistics, yielding within-assay variability coefficients ranging from 4.9% to 11%. A range of approximately 5 x 10(4)-1 x 10(7) cells could be detected. This method may be generally applicable to study surface binding of virtually any microbial species, while obviating the need of radioactive materials or specific antibodies for quantification, thus providing a procedure that is useful to both basic and clinical research.
Bosch
J A
JA
Department of Dental Basic Sciences, Section Oral Biochemistry, Vrije Universiteit, Academic Centre for Dentistry Amsterdam (ACTA), Van der Boechorststraat 7 1081 BT, Amsterdam, The Netherlands.
Veerman
E C I
EC
Turkenburg
M
M
Hartog
K
K
Bolscher
J G M
JG
Nieuw Amerongen
A V
AV
D001422:Bacterial Adhesion; D001665:Binding Sites; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes; D016480:Helicobacter pylori / Q000235:genetics / Q000472:pathogenicity; D006801:Humans; D012463:Saliva / Q000382:microbiology*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D034361:Streptococcus mitis / Q000235:genetics / Q000472:pathogenicity; D013298:Streptococcus sanguis / Q000235:genetics / Q000472:pathogenicity
D004269:DNA, Bacterial; D005456:Fluorescent Dyes