Glucosyltransferase as a Marker for Caries Activity

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。评估龋齿活动和发生新病变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，1951；Duchin and van Houte 1978；Beighton, 1991]，牙菌斑和唾液中酸性微生物的测定[Snyder, 1951；Grainger等人，1965年]，以及先前龋齿活动的评估[Grainger和Nikiforuk, 1960；Stamm, 1993;大白鲟,1997;鲍威尔,1998;梅塞尔集团,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病，所有这些都有缺点。葡萄糖基转移酶（Gff， E.C. 2.4.1.5）是迄今为止从变形链球菌中鉴定出的龋齿病因中唯一被证实的毒力因子[DeStoppelaar等人，1971；Hamada et al., 1984；Tanzer et al., 1985；Yamashita等，1993]。唾液中活性Gff水平与变形链球菌唾液种群相关[R611a等，1983；Scheie et al., 1987；[j]。鉴于gff已被证明是龋病发病机制中的毒力因子，并且存在于整个唾液中，因此探索开发一种使用该酶作为龋病标志物的测试似乎是明智的。为此，我们将通过直接酶测定法检测无龋儿童或早期儿童龋齿儿童的唾液中Gff（由突变链球菌和非突变链球菌引起）。我们还将在酶联免疫吸附试验中使用单克隆抗体测定受试者唾液中变形链球菌的GffB、GtfC和GtID的数量。然后，我们将尝试将测定的Gff活性（来自突变链球菌和非突变链球菌）与变形链球菌的GtfB、GffC和GffD浓度联系起来。最后，我们将确定我们是否可以将GtfB、GtfC和GffD的浓度以及唾液中Gff的总体测定活性与受试者目前的临床龋齿经历水平相关联。如果在这个第一步的横断面试点研究中成功地显示出总体Gff活性或唾液中个体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（与本应用分开），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的风险，并识别那些在空化甚至白斑形成之前活跃的龋齿。

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.