Detection of group 1 Trypanosoma brucei gambiense by loop-mediated isothermal amplification

Njiru, Z. K., Traub, R. J., Ouma, J. O., Enyaru, J. C. and Matovu, E. (2011) Detection of group 1 Trypanosoma brucei gambiense by loop-mediated isothermal amplification. Journal of Clinical Microbiology, 49 4: 1530-1536. doi:10.1128/JCM.01817-10

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Author Njiru, Z. K.
Traub, R. J.
Ouma, J. O.
Enyaru, J. C.
Matovu, E.
Title Detection of group 1 Trypanosoma brucei gambiense by loop-mediated isothermal amplification
Journal name Journal of Clinical Microbiology   Check publisher's open access policy
ISSN 0095-1137
Publication date 2011-04
Sub-type Article (original research)
DOI 10.1128/JCM.01817-10
Open Access Status File (Publisher version)
Volume 49
Issue 4
Start page 1530
End page 1536
Total pages 7
Place of publication Washington, DC, United States
Publisher American Society for Microbiology
Collection year 2012
Language eng
Abstract Trypanosoma brucei gambiense group 1 is the major causative agent of the Gambian human African trypanosomiasis (HAT). Accurate diagnosis of Gambian HAT is still challenged by lack of precise diagnostic methods, low and fluctuating parasitemia, and generally poor services in the areas of endemicity. In this study, we designed a rapid loop-mediated isothermal amplification (LAMP) test for T. b. gambiense based on the 3= end of the T. b. gambiense-specific glycoprotein (TgsGP) gene. The test is specific and amplifies DNA from T. b. gambiense isolates and clinical samples at 62°C within 40 min using a normal water bath. The analytical sensitivity of the TgsGP LAMP was equivalent to 10 trypanosomes/ml using purified DNA and ̃1 trypanosome/ ml using supernatant prepared from boiled blood, while those of classical PCR tests ranged from 10 to 103 trypanosomes/ml. There was 100% agreement in the detection of the LAMP product by real-time gel electrophoresis and the DNA-intercalating dye SYBR green I. The LAMP amplicons were unequivocally confirmed through sequencing and analysis of melting curves. The assay was able to amplify parasite DNA from native cerebrospinal fluid (CSF) and double-centrifuged supernatant prepared from boiled buffy coat and bone marrow aspirate. The robustness, superior sensitivity, and ability to inspect results visually through color change indicate the potential of TgsGP LAMP as a future point-of-care test.
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Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2012 Collection
School of Veterinary Science Publications
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Created: Thu, 13 Oct 2011, 15:38:01 EST by Rebecca Traub on behalf of School of Veterinary Science