Diagnosis of gambiense HAT

描述(申请人提供)：布氏冈比亚锥虫和罗德赛锥虫是人类非洲锥虫病(HAT或昏睡病)的病原体。如果不治疗，死于帽子是不可避免的。最近，HAT已经达到流行的程度，非洲每年有超过4万人死亡。虽然对有限药物治疗的抗药性可能是一个因素，但在患者通常可见的农村地区，在疾病的关键早期阶段未能通过显微镜观察血液涂片和/或脑脊液中的锥虫可能是治疗失败的唯一最重要的因素。虽然使用基于聚合酶链式反应的方法进行HAT的早期检测是可能的，但由于使用的往往是价格不菲的“热循环仪”，可能会在高温、潮湿和/或多尘的环境中不稳定地发挥作用，因此，在HAT流行地区，使用聚合酶链式反应作为一种实用的诊断方法是不可能的。我们提出了一种最新的技术，我们认为这将对检测寄生虫感染非常重要，特别是在现场环境中。这是一种被称为环介导的等温扩增(LAMP)的PCR的替代方法。这种反应在等温条件下以高特异性、高效率和高速度扩增DNA，并允许容易地目视阳性鉴定目标DNA。我们研制了针对冈比亚和罗德氏锥虫引起的检测帽的灯。我们认为，LAMP将为现场使用的PCR和其他检测方法提供一种更容易、可能更特异的替代方法。在这项应用中，我们建议验证基于LAMP的检测在安哥拉HAT临床诊断中的潜力，重点是第二阶段HAT。这项提案的具体目标的成功完成将为早期发现适用于非洲保健中心和实地的HAT提供新的诊断测试。非洲昏睡病是由非洲锥虫(一种原生动物寄生虫)引起的，每年在非洲导致4万多人死亡。这是因为在农村地区不存在诊断和分期昏睡病的良好测试。我们想开发一种新的、简单的检测方法，用于这种致命疾病的早期诊断。

DESCRIPTION (provided by applicant): Trypanosoma brucei gambiense and T. b. rhodesiense are pathogens responsible for human African trypanosomiasis (HAT or sleeping sickness). Death from HAT is inevitable if untreated. Recently, HAT has reached epidemic proportions with over 40,000 people dying every year in Africa. While resistance to the limited drug therapies may be a factor, in the rural areas where patients are typically seen, failure to microscopically observe trypanosomes in blood smears and/or CSF in the critical early stages of the disease is probably the single most important factor in failed treatment. While early detection of HAT using PCR-based methods is possible, the use of often non- affordable "thermal cyclers" that may work erratically at high ambient temperatures, humidity and/or dusty environments, negates PCR as a practical diagnostic method in HAT endemic areas. We propose a recent technology that we believe will be important for the detection of parasite infections, particularly in the field setting. It is an alternative to PCR called loop-mediated isothermal amplification (LAMP). This reaction amplifies DNA with high specificity, efficiency and speed under isothermal conditions and allows for easy visual positive identification of the target DNA. We have developed LAMP for the detection HAT caused by T. b. gambiense and T. b. rhodesiense. We propose that LAMP will provide an easier, potentially more specific, alternative to PCR and other detection methods for use in the field. In this application we propose to validate the potential of LAMP-based tests for the clinical diagnosis of HAT in Angola with an emphasis on Stage 2 HAT. Success in the completion of the Specific Aims of this proposal will provide novel diagnostic tests for the early detection of HAT applicable in African health care centers and in the field. African sleeping sickness, caused by African trypanosomes (a protozoan parasite), kills over 40,000 people each year in Africa. This is because good tests for diagnosis and staging of sleeping sickness in rural areas do not exist. We want to develop a new and easy test for early diagnosis of this deadly disease.