DIAGNOSIS OF ENTAMOEBA HISTOLYTICA INFECTION

据信，溶组织内阿米巴感染了5亿人 每年一次。其中约10%的人出现症状并 发展为结肠炎和肝脓肿，死亡率超过 每年有4万人。目前的诊断方法是利用 显微镜。然而，这种方法需要丰富的经验，是 繁琐，如果不在几分钟内执行，可能不准确 标本收集。它也不区分非致病性和非致病性 致病菌株，这是一个主要缺点，因为非致病菌株 不会引起疾病，感染了非致病菌株的人也不会 需要用刺激性的抗阿米巴药物治疗。在第一阶段，我们启动了 两种单抗的制备及鉴定研究 (MAb)为基础的ELISA，可检测生物体的粘附素。研究结果是 对粪便标本进行检查，并将结果与显微镜进行比较， 培养和酵母菌分析。此外，我们还启动了关于 在第二阶段，我们检测了血清和肝脓肿抽吸物中的粘附素 将继续对溶组织埃希氏菌试验和开发进行临床评估 510(K)向FDA提交的申请。我们将开发出一种基因 高效表达溶组织埃希氏菌粘附素体系的建立 保存大便的条件，并评估“用户友好”的量水尺 测试。我们将开发其他检测方法来检测粘附素 血清、尿液和肝脏抽吸物。我们将发展聚合酶链式反应方法 用于评估的其他辅助工具。临床试验将在 重点放在艾滋病毒感染者身上。将收集510(K)的数据 向食品和药物管理局提交的申请。我们将特别强调 在完成第一阶段开始的工作后，进行高度具体的测试 致病性组织裂解肠杆菌。我们的测试将被放大以用于商业 申请。第二阶段代表了强大的合作努力，以开发 并改进阿米巴病的诊断测试。 拟议的商业应用：检测内阿米巴的免疫分析 粪便标本中的溶组织性肺炎及其与病原菌的鉴别 非致病菌株比方法更准确、更可靠 当前使用的。这个项目的结果应该会导致 商业诊断测试的发展将具有重要意义 对世界各地的健康产业具有重要价值。

Entamoeba histolytica is believed to infect a half billion people annually. Approximately 10% of these individuals become symptomatic and develop colitis and liver abscess with a mortality rate of more than 40,000 persons each year. The current method of diagnosis utilizes microscopy. However, this method requires extensive experience, is tedious, and may not be accurate if not performed within minutes of specimen collection. It also does not distinguish nonpathogenic from pathogenic strains, a major disadvantage because nonpathogenic strains do not cause disease and persons infected with nonpathogenic strains do not require treatment with harsh antiamebic drugs. In Phase I, we initiated studies on the development and evaluation of two monoclonal antibody (MAb)-based ELISAs that detect the adhesin of the organism. Studies were performed on stool specimens and results were compared with microscopy, culture, and zymodeme analysis. In addition, we initiated studies on the detection of adhesin in sera and liver abscess aspirates, In Phase II, we will continue clinical evaluations of the E. histolytica Test and develop 510(k) applications for submission to the FDA. We will develop a genetic system for overexpressing the E. histolytica adhesin, develop assay conditions for preserved stools, and evaluate "user-friendly" dipstick tests. We will develop alternative assay formats to detect adhesin in sera, urine, and liver aspirates. We will develop PCR methods as additional aids for evaluation. Clinical trials will be performed with emphasis on HIV-infected individuals. Data will be collected for 510(k) applications for submission to the FDA. Special emphasis will be placed on completing the work started in Phase I on tests highly specific for pathogenic E. histolytica. Our tests will be scaled up for commercial application. Phase II represents a strong collaborative effort to develop and improve diagnostic tests for amebiasis. PROPOSED COMMERCIAL APPLICATION: Immunoassays that detect Entamoeba histolytica in stool specimens and that distinguish pathogenic from nonpathogenic strains are more accurate and dependable than methods currently used. The results of this project should lead to the development of commercial diagnostic tests that will be of significant value to the health industry around the world.