Loop-mediated Isothermal Amplification (LAMP) test for detection of Trypanosoma evansi strain B

Njiru, Zablon K., Ouma, Johnson O., Enyaru, John C. and Dargantes, Alan P. (2010) Loop-mediated Isothermal Amplification (LAMP) test for detection of Trypanosoma evansi strain B. Experimental Parasitology, 125 3: 196-201. doi:10.1016/j.exppara.2010.01.017


Author Njiru, Zablon K.
Ouma, Johnson O.
Enyaru, John C.
Dargantes, Alan P.
Title Loop-mediated Isothermal Amplification (LAMP) test for detection of Trypanosoma evansi strain B
Formatted title
Loop-mediated Isothermal Amplification (LAMP) test for detection of Trypanosoma evansi strain B
Journal name Experimental Parasitology   Check publisher's open access policy
ISSN 0014-4894
1090-2449
Publication date 2010-07
Sub-type Article (original research)
DOI 10.1016/j.exppara.2010.01.017
Volume 125
Issue 3
Start page 196
End page 201
Total pages 6
Place of publication Maryland Heights, MO, United States
Publisher Academic Press
Language eng
Formatted abstract
Camel Trypanosomiasis (Surra) is mainly caused by Trypanosoma evansi strains that express variable surface glycoprotein (VSG) RoTat 1.2. However, in Kenya a second causative strain that does not express RoTat 1.2 VSG (T. evansi type B) has been identified. The prevalence of T. evansi type B largely remains unknown due to inadequate diagnostic assay. This work reports the development of a sensitive and specific diagnostic assay capable of detecting T. evansi type B based on the strategy of Loop-mediated Isothermal Amplification (LAMP) of DNA. The test is rapid and amplification is achieved within 20–25 min at 63 °C using a real time PCR machine. Restriction enzyme AluI digestion of the amplicon gave the predicted 83 bp and 89 bp sized bands and the LAMP product melt curves showed consistent melting temperature (Tm) of ∼89 °C. The assay analytical sensitivity is ∼0.1 tryps/ml while that of classical PCR test targeting the same gene is ∼10 tryps/ml. There was a 100% agreement in detection of the LAMP amplification product in real time, gel electrophoresis, on addition of SYBR Green I, and when using chromatographic Lateral Flow Dipstick (LFD) format. The use of the LAMP test revealed nine more T. evansi type B DNA samples that were not initially detected through PCR. The robustness and higher sensitivity of the T. evansi type B LAMP assay coupled with the visual detection of the amplification product indicate that the technique has strong potential as a point-of-use test in surra endemic areas.
Keyword Loop-mediated Isothermal Amplification (LAMP)
Trypanosoma evansi type B
Diagnosis
Lateral Flow Dipstick
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: ERA 2012 Admin Only
School of Veterinary Science Publications
 
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Created: Tue, 08 Nov 2011, 11:16:22 EST by Dr Zablon Njiru on behalf of School of Veterinary Science