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.

Public Health Relevance

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.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI082695-01
Application #
7666449
Study Section
Special Emphasis Panel (ZAI1-GSM-M (J2))
Program Officer
Rogers, Martin J
Project Start
2009-04-03
Project End
2013-03-31
Budget Start
2009-04-03
Budget End
2010-03-31
Support Year
1
Fiscal Year
2009
Total Cost
$426,762
Indirect Cost
Name
Johns Hopkins University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Nikolskaia, Olga V; Thekisoe, Oriel M M; Dumler, J Stephen et al. (2017) Loop-Mediated Isothermal Amplification for Detection of the 5.8S Ribosomal Ribonucleic Acid Internal Transcribed Spacer 2 Gene Found in Trypanosoma brucei gambiense. Am J Trop Med Hyg 96:275-279
Burbulis, Ian E; Yamaguchi, Kumiko; Nikolskaia, Olga V et al. (2015) Detection of pathogen-specific antibodies by loop-mediated isothermal amplification. Clin Vaccine Immunol 22:374-80
Sumpio, Brandon J; Chitragari, Gautham; Moriguchi, Takeshi et al. (2015) African Trypanosome-Induced Blood-Brain Barrier Dysfunction under Shear Stress May Not Require ERK Activation. Int J Angiol 24:41-6
Frevert, Ute; Movila, Alexandru; Nikolskaia, Olga V et al. (2012) Early invasion of brain parenchyma by African trypanosomes. PLoS One 7:e43913
Grab, Dennis J; Nikolskaia, Olga V; Inoue, Noboru et al. (2011) Using detergent to enhance detection sensitivity of African trypanosomes in human CSF and blood by loop-mediated isothermal amplification (LAMP). PLoS Negl Trop Dis 5:e1249
Grab, Dennis J; Garcia-Garcia, Jose C; Nikolskaia, Olga V et al. (2009) Protease activated receptor signaling is required for African trypanosome traversal of human brain microvascular endothelial cells. PLoS Negl Trop Dis 3:e479