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

Experimental Parasitology
Zablon K NjiruAlan P Dargantes

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 20-25min at 63 degrees C using a real time PCR machine. Restriction enzyme AluI digestion of the amplicon gave the predicted 83bp and 89bp sized bands and the LAMP product melt curves showed consistent melting temperature (T(m)) of approximately 89 degrees C. The assay analytical sensitivity is approximately 0.1tryps/ml while that of classical PCR test targeting the same gene is approximately 10tryps/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 reveal...Continue Reading

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Citations

Nov 10, 2010·PloS One·Naomi W LucchiVenkatachalam Udhayakumar
Jun 11, 2011·Trends in Parasitology·Sally L Wastling, Susan C Welburn
Aug 21, 2015·Parasitology Research·Thu-Thuy NguyenNoboru Inoue
May 29, 2013·Asian Pacific Journal of Tropical Medicine·Windell L Rivera, Vanissa A Ong
Jan 25, 2018·Parasitology·Christine M KamidiGrace Murilla
Apr 2, 2016·PLoS Neglected Tropical Diseases·Hadush BirhanuNick Van Reet

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