Glucosyltransferase as a Marker for Caries Activity

葡萄糖基转移酶作为龋齿活动的标志物

• 批准号: 6915780
• 负责人: WILLIAM H BOWEN
• 金额: $ 15.6万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6915780
• 关键词: Streptococcus; Streptococcus mutans; biomarker; clinical research; dental caries; disease /disorder proneness /risk; enzyme activity; enzyme linked immunosorbent assay; glucans; hexosyltransferase; human subject; monoclonal antibody; pellicle; preschool child (1-5); protein quantitation /detection; saliva

DESCRIPTION: Dental caries involves the interaction of host molecules, bacterial components, dietary substrate and time. Tests for assessing caries activity, and the likelihood of developing new lesions include determination of level of infection by using salivary mutans streptococci and lactobacilli counts [Rogosa, et al., 1951; Duchin and van Houte 1978; Beighton, 1991], determination of aciduric organisms in plaque and saliva [Snyder, 1951; Grainger et al., 1965], and assessment of prior caries activity [Grainger and Nikiforuk, 1960; Stamm, 1993; Hausen, 1997; Powell, 1998; Messer, 2000]. There is a need for a simple, reliable and reproducible test to determine caries in affected individuals, all of which have shortcomings. Glucosyltransferases (Gff, E.C. 2.4.1.5) are the only proven virulence factors in the etiology of Dental caries identified from Streptococcus mutans thus far [DeStoppelaar et al., 1971; Hamada et al., 1984; Tanzer et al., 1985; Yamashita et al., 1993]. Levels of active Gff in saliva correlate with salivary populations of S. mutans [R611a et al., 1983; Scheie et al., 1987; Vacca-Smith et al., 1996b]. Given that Gffs are proven virulence factors in the pathogenesis of Dental caries and are present in whole saliva, it appears prudent to explore the development of a test using the enzyme(s) as a marker(s) for caries. To this end we will test saliva from either caries free children, or children with Early Childhood Caries, for Gff (contributed by both mutans and non-mutans streptococci) by direct enzyme assay. We will also determine the quantity(ies) of GffB, GtfC, and GtID of S. mutans in the subjects' saliva using monoclonal antibodies in an enzyme-linked immunosorbent assay. We will then attempt to correlate the assayed activity of Gff (from both mutans and non-mutan streptococci) with the concentrations of GtfB, GffC and GffD of S. mutans. Finally, we will determine whether we can correlate both the concentrations of GtfB, GtfC, and GffD, and the overall assayed Gff activity in saliva, with the current levels of clinical caries experience of the subjects. If successful in showing, in this first-step cross-sectional pilot study, a correlation between overall Gff activity, or level of individual Gffs in saliva, with caries prevalence, then the way might be open to explore in the future, in a second, longitudinal study, (separate from this application) whether Gtf in saliva can be used as a predictor of future caries development. Our eventual goal (in a future application) is to determine caries risk well before the onset of lesions and to identify those who are caries active short of cavitation or even white spot formation.

描述：龋齿涉及宿主分子，细菌成分，饮食基质和时间的相互作用。用于评估龋齿活性和发展新病变的可能性的测试包括通过使用唾液变形链球菌和乳酸杆菌计数来确定感染水平[Rogosa等人，1951; Duchin和货车Houte，1978; Beighton，1991]，牙菌斑和唾液中酸尿微生物的测定[Snyder，1951; Grainger等人，1965]，以及评估既往龋齿活动性[Grainger和Nikiforuk，1960; Stamm，1993;豪森，1997; Powell，1998; Messer，2000]。需要一种简单、可靠和可重复的测试来确定受影响个体的龋齿，所有这些都有缺点。葡萄糖基转移酶（Gff，EC 2.4.1.5）是迄今为止从变形链球菌中鉴定出的龋齿病因学中唯一经证实的毒力因子[DeStoppelaar等人，1971; Hamada等人，1984; Tanzer等人，1985; Yamashita等人，1993年]。唾液中活性Gff的水平与唾液中链球菌的数量相关。变异株[R611 a等，1983; Scheie等人，1987; Vacca-Smith等人，1996年b]。鉴于Gff是龋齿发病机制中已证实的毒力因子，并且存在于整个唾液中，探索使用酶作为龋齿标记物的测试开发似乎是谨慎的。为此，我们将通过直接酶测定法测试无龋儿童或患有幼儿龋的儿童的唾液中的Gff（由变形链球菌和非变形链球菌贡献）。我们还将确定S的GffB、GtfC和GtID的数量。在酶联免疫吸附测定中使用单克隆抗体检测受试者唾液中的变形杆菌。然后，我们将尝试将Gff（来自变链菌和非变链菌）的测定活性与S的GtfB、GffC和GffD浓度相关联。变异人最后，我们将确定我们是否可以将唾液中GtfB、GtfC和GffD的浓度以及总体测定的Gff活性与受试者的当前临床龋齿经历水平相关联。如果在第一步的横断面初步研究中成功地显示出总体Gff活性或唾液中单个Gff水平与龋齿患病率之间的相关性，那么在未来的第二次纵向研究中（与本申请分开），可能会探索唾液中的Gtf是否可以用作未来龋齿发展的预测因子。我们的最终目标（在未来的应用中）是在病变发生之前确定龋齿风险，并识别出那些龋齿活跃的人，这些人没有空洞甚至没有白色斑点形成。