Use of GtfB as a Diagnostic for Caries Activity

使用 GTfB 作为龋齿活动的诊断

• 批准号: 7482074
• 负责人: John Linforth Daiss
• 金额: $ 18.6万
• 依托单位: FIRST WAVE TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7482074
• 关键词: Acids; Address; Adsorption; Antibodies; Area Under Curve; Bacteria; Biological Assay; Biological Markers; Child; Childhood; Cleaved cell; Clinical; Clinical Trials; Cross-Sectional Studies; Data; Dental; Dental Enamel; Dental Offices; Dental caries; Dentists; Dentition; Detection; Development; Diagnostic; Diagnostic tests; Dietary Sucrose; Disease; Disease Progression; Early Intervention; Energy-Generating Resources; Enzymes; Epidemic; Epitopes; Freezing; Frequencies; Fructose; Generations; Glucans; Glucose; Goals; Human; Immunoassay; Immunoblotting; Immunoglobulin A; Immunoglobulin G; Intervention; Laboratories; Lesion; Longitudinal Studies; Low income; Measurement; Measures; Methods; Microbial Biofilms; Monoclonal Antibodies; Monosaccharides; Morbidity - disease rate; Office Visits; Pathogenesis; Pathologic; Patients; Performance; Pilot Projects; Population; Populations at Risk; Predictive Value; Probability; Public Health; Receiver Operator Characteristics; Recovery; Reporting; Risk; Role; Running; Rural Population; Saliva; Salivary; Sampling; Schools; Site; Source; Streptococcus mutans; Surgeon; Testing; Time; Tooth structure; Trauma; United States; Validation; Variant; White Spots; base; clinical efficacy; cost; diagnostic accuracy; early childhood; glucosyltransferase B; glucosyltransferase D; glycosyltransferase; instrument; pathogen; pediatrician; prevent; prototype; repaired; success; tool; tooth surface

DESCRIPTION (provided by applicant): The over-arching objective of this application is the creation of a simple, rapid, inexpensive diagnostic test that can be used during an office visit to detect caries activity before overt carious lesions are apparent so that interventions can be made before permanent damage to the dentition occurs. The specific objective of this application is the clinical validation of an immunoassay measuring GtfB as a saliva-based marker for caries activity in children from a population where Early Childhood Caries (ECC) is prevalent. This method is predicated on an intimate understanding of the pathogenesis of Dental caries. Three glycosyltransferases (GtfB, GtfC and GtfD) are secreted by S. mutans when it colonizes the tooth surface. These enzymes cleave dietary sucrose into its constituent monosaccharides, glucose which is polymerized into soluble and insoluble glucans that create the tightly adherent, bacteria-rich biofilm that characterizes caries, and fructose which is utilized as an energy source and yields enamel-dissolving acids. The central role of Gtfs in disease progression makes them plausible markers for active caries. In a recent cross-sectional clinical trial, the Bowen laboratory distinguished children with ECC from disease-free children with about 80% diagnostic accuracy by measuring the levels of GtfB in their saliva. With such a promising start, two issues become paramount: 1) whether the GtfB immunoassay will be an effective tool for identifying caries-active children prospectively; and 2) ways by which the clinical performance of the assay can be optimized while the method is made useable in an office setting. This application addresses exactly these questions. The focus of Specific Aim 1 is a 100 patient clinical trial to determine whether GtfB measurement can identify children who have the disease Dental caries before they have clinically detectable carious lesions. Clinically caries-free children 24-60 months old will be examined for white spot lesions or frank cavitation every three months for a year. Saliva samples will be taken at the same time and GtfB measured. The clinical question will be: It is not clear whether elevated salivary GtfB values predict the emergence of detectable carious lesions 3, 6 or 9 months later. Specific Aims 2 and 3 address the creation of additional anti-GtfB monoclonal antibodies and alternative immunoassay formats so we can ultimately make a saliva-based, rapid, inexpensive, instrument-free diagnostic test for incipient Dental caries. Public Health Significance: Use of GtfB as a Diagnostic for Caries Activity. Dental caries remains a major source of morbidity and expense. In the United States alone, $48 billion are spent each year repairing the consequences of this completely preventable disease. A particularly severe form, Early Childhood Caries (ECC) is common in impoverished urban and rural populations creating what the Report of the Surgeon General called a silent epidemic. Among the tools needed for reducing the burden of Dental caries is a simple, inexpensive, in-office, diagnostic test that can identify children with active caries so that interventions can be made before their teeth become irreversibly damaged. The diagnostic must be simple and rapid enough that it can be used in a pediatric or Dental office. The first goal is to validate a saliva-borne, pathogen-derived marker that is the proposed analyte in such a longitudinal test.

描述（由申请人提供）：本申请的主要目的是创建一种简单、快速、廉价的诊断测试，该测试可在门诊期间使用，以在明显的龋齿损伤明显之前检测龋齿活动，从而可以在牙列发生永久性损伤之前进行干预。本申请的具体目的是对一种免疫测定法进行临床验证，该免疫测定法测量GtfB作为儿童早期龋齿（ECC）流行人群中龋齿活动的唾液标志物。这种方法是基于对龋齿发病机制的深入了解。葡萄糖基转移酶（GtfB、GtfC和GtfD）是葡萄糖基转移酶的主要组成部分。当它定植在牙齿表面时会发生变异。这些酶将膳食蔗糖裂解成其组成单糖，葡萄糖聚合成可溶性和不溶性葡聚糖，产生紧密粘附的、富含细菌的生物膜，其特征在于龋齿，以及果糖，其用作能源并产生溶解釉质的酸。Gtfs在疾病进展中的核心作用使其成为活动性龋齿的合理标记。在最近的一项横断面临床试验中，Bowen实验室通过测量唾液中GtfB的水平，以约80%的诊断准确率区分ECC儿童和无病儿童。有了这样一个有希望的开始，两个问题变得至关重要：1）GtfB免疫测定是否将是一个有效的工具，用于识别龋齿活跃的儿童前瞻性;和2）的方法，通过该方法可以优化的临床性能，同时该方法是在办公室设置可用。这个应用程序正好解决了这些问题。具体目标1的重点是一项100名患者的临床试验，以确定GtfB测量是否可以在临床上可检测到龋齿病变之前识别患有龋齿的儿童。临床上无龋儿童24-60个月将检查白色斑点病变或坦率的空洞每三个月一年。同时采集唾液样本并测量GtfB。临床问题将是：目前尚不清楚唾液GtfB值升高是否预测3、6或9个月后可检测到的龋损的出现。具体目标2和3解决了额外的抗GtfB单克隆抗体和替代免疫测定形式的创建，因此我们最终可以针对早期龋齿进行基于唾液的、快速的、廉价的、无仪器的诊断测试。公共卫生意义：使用GtfB作为龋齿活动的诊断。龋齿仍然是发病率和费用的主要来源。仅在美国，每年就花费480亿美元来修复这种完全可以预防的疾病的后果。一种特别严重的形式，幼儿龋齿（ECC）在贫困的城市和农村人口中很常见，创造了卫生部长报告中所说的无声流行病。在减少龋齿负担所需的工具中，有一种简单、廉价的诊所内诊断测试，可以识别患有活动性龋齿的儿童，以便在他们的牙齿受到不可逆的损害之前进行干预。诊断必须足够简单和快速，以便在儿科或牙科诊所使用。第一个目标是验证唾液传播的病原体衍生的标记物，该标记物是在这样的纵向测试中提出的分析物。