TY - JOUR
T1 - In situ and non-invasive detection of specific bacterial species in oral biofilms using fluorescently labeled monoclonal antibodies
AU - Gu, Fang
AU - Lux, Renate
AU - Du-Thumm, Laurence
AU - Stokes, Ivy
AU - Kreth, Jens
AU - Anderson, Maxwell H.
AU - Wong, David T.
AU - Wolinsky, Lawrence
AU - Sullivan, Richard
AU - Shi, Wenyuan
N1 - Funding Information:
We thank Fengxia Qi for helpful discussions. This work was supported by grants from the Colgate-Palmolive Company, C3 Scientific Corporation, Washington Dental Service and NIH DE15018.
PY - 2005/8
Y1 - 2005/8
N2 - Noninvasive in situ detection of suspected cariogenic bacterial species within dental biofilms could facilitate monitoring of the dynamic change of oral microbial flora and assist in the assessment of the treatment efficacy of therapeutic agents. In this study, we explore the possibility to use three well-characterized monoclonal antibodies (MAbs) against Streptococcus mutans, Actinomyces naeslundii, and Lactobacillus casei to identify these three important members of the oral microbial community in the complex environment of oral biofilms. These MAbs, which were conjugated to different fluorescent labels and visualized with confocal laser scanning microscopy (CLSM), proved to be an useful tool to identify the three species of interest (S. mutans, A. naeslundii, and L. casei) under various experimental conditions including in vitro and in vivo derived oral biofilms. Manifold addition of the MAbs on consecutive days did not alter the biofilm structure thus allowing monitoring of the same biofilm over extended time periods. Using this MAb-based method the effect of sucrose challenge on the biofilm composition and the distribution of S. mutans, A. naeslundii, and L. casei were examined. S. mutans was found to be the predominant species under the various biofilm conditions tested. These studies indicate that MAbs based bacterial detection with CLSM is a versatile tool which permits new insights into the ecology of oral biofilm development.
AB - Noninvasive in situ detection of suspected cariogenic bacterial species within dental biofilms could facilitate monitoring of the dynamic change of oral microbial flora and assist in the assessment of the treatment efficacy of therapeutic agents. In this study, we explore the possibility to use three well-characterized monoclonal antibodies (MAbs) against Streptococcus mutans, Actinomyces naeslundii, and Lactobacillus casei to identify these three important members of the oral microbial community in the complex environment of oral biofilms. These MAbs, which were conjugated to different fluorescent labels and visualized with confocal laser scanning microscopy (CLSM), proved to be an useful tool to identify the three species of interest (S. mutans, A. naeslundii, and L. casei) under various experimental conditions including in vitro and in vivo derived oral biofilms. Manifold addition of the MAbs on consecutive days did not alter the biofilm structure thus allowing monitoring of the same biofilm over extended time periods. Using this MAb-based method the effect of sucrose challenge on the biofilm composition and the distribution of S. mutans, A. naeslundii, and L. casei were examined. S. mutans was found to be the predominant species under the various biofilm conditions tested. These studies indicate that MAbs based bacterial detection with CLSM is a versatile tool which permits new insights into the ecology of oral biofilm development.
KW - Actinomyces naeslundii
KW - Lactobacillus casei
KW - Monoclonal antibodies
KW - Oral biofilms
KW - Streptococcus mutans
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U2 - 10.1016/j.mimet.2005.02.013
DO - 10.1016/j.mimet.2005.02.013
M3 - Article
C2 - 15935497
AN - SCOPUS:22244476157
SN - 0167-7012
VL - 62
SP - 145
EP - 160
JO - Journal of Microbiological Methods
JF - Journal of Microbiological Methods
IS - 2
ER -