PCR-BASED DIAGNOSIS OF HELICOBACTER PYLORI

基于 PCR 的幽门螺杆菌诊断

• 批准号: 2656646
• 负责人: TERESA A LEHMAN
• 金额: $ 9.91万
• 依托单位: BIOSERVE BIOTECHNOLOGIES, LTD
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 1998-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2656646
• 关键词: African American; Helicobacter; Hispanic Americans; biomarker; clinical chemistry; clinical research; communicable disease diagnosis; diagnosis design /evaluation; enteric bacteria; fluorimetry; globin; human subject; noninvasive diagnosis; polymerase chain reaction; saliva; urease

Helicobacter pylori (H. pylori) infection is the most frequent bacterial infection worldwide. H. pylori is a major etiological agent in the development of gastritis, gastric ulcer, and gastric cancer. To develop an ultrasensitive, non-invasive screening method for the presence of H. pylori in saliva and other clinical samples, a PCR coamplification assay of urease and beta-globin, followed by fluorometric detection of modified products, is proposed. If the urease gene of H. pylori is present, urease-specific biotin and HEX primers will be physically associated in the double stranded PCR product. Incubation of PCR products in streptavidin coated plates will allow binding of biotinylated primers to streptavidin. The presence of the H. pylori specific PCR product will be evaluated by the detection of fluorescence of the HEX primers. If the screening assay is successful in detecting extremely low levels of H. pylori in the clinical samples, it could be used as a biomarker for early indication of gastric diseases. The commercial application of a screening kit for H. pylori would be tremendous. Proposed commercial applications: This research project for the development of a diagnostic test for the presence of H. pylori could be more sensitive and specific than current methods. The commercial application is tremendous since this is the most frequent bacterial infection worldwide. The methodology is amenable to large scale screening.

幽门螺杆菌 (H. pylori) 感染是最常见的细菌 感染全球。 幽门螺杆菌是一种主要的病原体 胃炎、胃溃疡和胃癌的发展。 发展 一种超灵敏、非侵入性的幽门螺杆菌筛查方法。 唾液和其他临床样本中的幽门螺杆菌，PCR 辅助扩增测定 脲酶和β-珠蛋白，然后用荧光检测修饰 产品，建议。 如果存在幽门螺杆菌脲酶基因， 脲酶特异性生物素和 HEX 引物将在物理上关联 双链PCR产物。 PCR 产物的孵育 链霉亲和素包被的板将允许生物素化引物结合 链霉亲和素。 幽门螺杆菌特异性 PCR 产物的存在将是 通过检测 HEX 引物的荧光来评估。 如果 筛选试验成功地检测出极低水平的幽门螺杆菌。 临床样本中的幽门螺杆菌可作为早期诊断的生物标志物 胃病的征兆。 筛选的商业应用 幽门螺杆菌试剂盒将是巨大的。 拟议的商业应用： 该研究项目旨在开发一种诊断测试 幽门螺杆菌的存在可能比目前的检测更灵敏、更特异 方法。 商业应用是巨大的，因为这是最 全球细菌感染频繁发生。 该方法论适合 大规模筛选。