Loop Mediated Isothermal Amplification for Detection of Trypanosoma brucei gambiense in Urine and Saliva Samples in Nonhuman Primate Model.
Human African trypanosomiasis (HAT) is a vector-borne parasitic zoonotic disease. The disease caused by Trypanosoma brucei gambiense is the most prevalent in Africa. Early diagnosis is hampered by lack of sensitive diagnostic techniques. This study explored the potential of loop mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) in the detection of T. b. gambiense infection in a vervet monkey HAT model. Six vervet monkeys were experimentally infected with T. b. gambiense IL3253 and monitored for 180 days after infection. Parasitaemia was scored daily. Blood, cerebrospinal fluid (CSF), saliva, and urine samples were collected weekly. PCR and LAMP were performed on serum, CSF, saliva, and urine samples. The detection by LAMP was significantly higher than that of parasitological methods and PCR in all the samples. The performance of LAMP varied between the samples and was better in serum followed by saliva and then urine samples. In the saliva samples, LAMP had 100% detection between 21 and 77 dpi, whereas in urine the detection it was slightly lower, but there was over 80% detection between 28 and 91 dpi. However, LAMP could not detect trypanosomes in either saliva or urine after 140 and 126 dpi, respectively. The findings of this study emphasize the importance of LAMP in diagnosis of HAT using saliva and urine samples.
Animal Science Department, Institute of Primate Research (IPR), P.O. Box 24481, Karen, Nairobi 00502, Kenya.
Maina
M
Ngotho
Animal Health and Production Department, College of Agriculture and Natural Resources, Jomo Kenyatta University of Agriculture and Technology (JKUAT), P.O. Box 62000, Nairobi 00200, Kenya.
John Maina
JM
Kagira
Biochemistry Department, College of Health Sciences, Jomo Kenyatta University of Agriculture and Technology (JKUAT), P.O. Box 62000, Nairobi 00200, Kenya.
Beatrice Muthoni
BM
Gachie
Public Health Department, College of Health Sciences, Jomo Kenyatta University of Agriculture and Technology (JKUAT), P.O. Box 62000, Nairobi 00200, > Kenya.
Simon Muturi
SM
Karanja
Biochemistry Department, College of Health Sciences, Jomo Kenyatta University of Agriculture and Technology (JKUAT), P.O. Box 62000, Nairobi 00200, Kenya.
Maxwell Wambua
MW
Waema
Animal Science Department, Institute of Primate Research (IPR), P.O. Box 24481, Karen, Nairobi 00502, Kenya.
Dawn Nyawira
DN
Maranga
Biochemistry Department, College of Health Sciences, Jomo Kenyatta University of Agriculture and Technology (JKUAT), P.O. Box 62000, Nairobi 00200, Kenya.
Naomi Wangari
NW
Maina
D000818:Animals; D002522:Chlorocebus aethiops; D016054:DNA, Protozoan / Q000032:analysis* / Q000302:isolation & purification / Q000652:urine*; D004195:Disease Models, Animal; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010274:Parasitology; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D012463:Saliva / Q000469:parasitology*; D014347:Trypanosoma brucei gambiense / Q000235:genetics* / Q000302:isolation & purification; D014353:Trypanosomiasis, African / Q000175:diagnosis* / Q000469:parasitology
D016054:DNA, Protozoan