The use of Loop-mediated Isothermal Amplification (LAMP) to detect the re-emerging Human African Trypanosomiasis (HAT) in the Luangwa and Zambezi valleys

Abstract

BackgroundLoop-mediated isothermal amplification (LAMP) is a novel strategy which amplifies DNA with high sensitivity and rapidity under isothermal conditions. In the present study, the performance of the repetitive insertion mobile element (RIME)-LAMP and human serum resistance-associated gene (SRA)-LAMP assays were evaluated using clinical specimens obtained from four male patients from Luangwa and Zambezi valleys in Zambia and Zimbabwe, respectively.FindingsThe cases reported in this preliminary communication were all first diagnosed by microscopy, through passive surveillance, and confirmed by both RIME-LAMP and SRA-LAMP. A good correlation between microscopy and LAMP was observed and contributed to staging and successful treatment of patient. RIME-LAMP and SRA-LAMP complimented each other well in all the cases.ConclusionsBoth RIME-LAMP and SRA-LAMP were able to detect Trypanosoma brucei rhodesiense DNA in patient blood and CSF and hence confirmed HAT in the parasitaemic patients. Our study indicates that the LAMP technique is a potential tool for HAT diagnosis, staging and may be useful for making therapeutic decisions. However, no statistically significant conclusion may be drawn due to the limited sample size used in the present study. It is thus imperative to conduct a detailed study to further evaluate the potential of LAMP as a bedside diagnostic test for HAT.

Prevalence and species distribution of caprine trypanosomosis in Sinazongwe and Kalomo districts of Zambia

“…The resultant DNA concentrate was stored at −20 • C until subjected to molecular analysis. Details of the protocol used are described elsewhere (Ahmadu et al, 2002;Namangala et al, 2012).…”

“…A LAMP reaction of 25 l total reaction volume was performed using a Loopamp DNA Amplification Kit (Eiken Chemical, Tochigi, Japan) and the extracted DNA as template, as described by Thekisoe et al (2007) and Namangala et al (2012). Recently described primers for amplification of CON2 18s rRNA (Thekisoe et al, 2007); and RIME, SRA and Satellite DNA (Njiru et al, 2008a(Njiru et al, ,2008b(Njiru et al, , 2011 were used for Trypanasoma congolense-LAMP, repetitive insertion mobile element (RIME)-LAMP, human serum resistance associated gene (SRA)-LAMP, and Trypanasoma vivax-LAMP trypanosome species detection and characterization (Table 1).…”

Preliminary Investigation of Trypanosomosis in Exotic Dog Breeds from Zambia's Luangwa and Zambezi Valleys Using LAMP

“…rhodesiense. In view of the sporadic cases of re-emerging human African trypanosomiasis (HAT) being reported in the old foci, 6 the finding of SRA gene in trypanosomes isolated from some dogs in this communication deserves further investigations. Considering the close relationship between dogs and humans, dogs may be a potential source of HAT, which may result when tsetse flies bite humans after taking a blood meal from a trypanosome-infected dog.…”

“…6 However, microscopy, the current gold standard diagnostic test for trypanosomosis, suffers from low sensitivity. Here, we evaluated the performance of LAMP *Address correspondence to Boniface Namangala, University of Zambia, School of Veterinary Medicine, Department of Paraclinical Studies, P.O.…”

“…A LAMP reaction of 25 mL was performed using a Loopamp DNA Amplification Kit (Eiken Chemical, Tochigi, Japan) and the extracted DNA as template. 6,7 This study made use of the recently designed primers specifically targeting the 18S rRNA gene of T. congolense (CON2-LAMP), 7 the repetitive insertion mobile element (RIME) gene of the Trypanozoon subgenus group (RIME-LAMP) 8 and the human serum resistance-associated (SRA) gene uniquely expressed by T.b. rhodesiense (SRA-LAMP), 9 respectively.…”