Diagnosis of gambiense HAT

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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.