Glucosyltransferase as a Marker for Caries Activity

描述：龋病涉及宿主分子、细菌成分、饮食底物和时间的相互作用。用于评估龋病活动性和发展新病变的可能性的测试包括通过唾液变形链球菌和乳杆菌计数来确定感染水平[Rogosa，等人，1951；Duchin和van Houte 1978；Beighton，1991]，牙菌斑和唾液中的耐酸性微生物的测定[Snyder，1951；Grainger等人，1965]，以及对以前的龋病活动的评估[Grainger和Nikiforuk，1960；Stamm，1993；Hausen，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人的龋齿，所有这些测试都有缺点。糖基转移酶(GFF，E.C.2.4.1.5)是迄今为止从变形链球菌中发现的唯一已证实的龋病病因学毒力因子[DeStoppelaar等人，1971；Hamada等人，1984；Tanzer等人，1985；Yamashita等人，1993]。唾液中活跃的GFF水平与变形链球菌的唾液种群相关[R611a等人，1983；Scheie等人，1987；Vacca-Smith等人，1996 b]。鉴于Gff是已被证实的龋病发病机制中的致病因子，并且存在于整个唾液中，因此探索一种使用酶(S)作为龋病标志物(S)的检测方法似乎是谨慎的。为此，我们将通过直接酶分析检测无龋儿童或有早期龋齿儿童的唾液中的GFF(由变形链球菌和非变形链球菌贡献)。我们还将在酶联免疫吸附试验中使用单抗测定受试者唾液中变形链球菌的gffB、gtfc和gtID的数量。然后，我们将尝试将检测到的GFF活性(来自变形链球菌和非突变链球菌)与变形链球菌的GTFB、GFFC和GffD的浓度相关联。最后，我们将确定是否可以将唾液中GTFB、GTFc和GffD的浓度以及所检测的整体GFF活性与受试者当前的临床龋病水平相关联。如果在这项第一步的横断面先导性研究中，成功地显示了总体GFF活性或唾液中单个GFF水平与龋齿患病率之间的相关性，那么在未来的第二项纵向研究中(与这项应用分开)，可能还可以探索唾液中的GTF是否可以用作未来龋病发展的预测因子。我们的最终目标(在未来的应用中)是在病变发生之前很久就确定龋齿风险，并识别那些龋齿活跃、空洞或甚至白斑形成的人。

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.