TY - JOUR
T1 - A rapid solid-phase fluorimetric assay for measuring bacterial adherence, using DNA-binding stains
AU - Bosch, J.A.
AU - Veerman, E.C.I.
AU - Turkenburg, M.
AU - Hartog, K.
AU - Bolscher, J.G.M.
AU - Nieuw Amerongen, A.V.
PY - 2003
Y1 - 2003
N2 - 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 5×104–1×107 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.
AB - 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 5×104–1×107 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.
U2 - https://doi.org/10.1016/S0167-7012(02)00220-8
DO - https://doi.org/10.1016/S0167-7012(02)00220-8
M3 - Article
SN - 0167-7012
VL - 53
SP - 51
EP - 56
JO - Journal of Microbiological Methods
JF - Journal of Microbiological Methods
IS - 1
ER -