Loop Mediated Isothermal Amplification for Detection ofTrypanosoma brucei gambiensein Urine and Saliva Samples in Nonhuman Primate Model

Abstract

Human African trypanosomiasis (HAT) is a vector-borne parasitic zoonotic disease. The disease caused byTrypanosoma brucei gambienseis 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 ofT. b. gambienseinfection in a vervet monkey HAT model. Six vervet monkeys were experimentally infected withT. b. gambienseIL3253 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.