Stability of Loop-Mediated Isothermal Amplification (LAMP) Reagents and its Amplification Efficiency on Crude Trypanosome DNA Templates

• DOI: 10.1292/jvms.71.471
• Publication Date: 2009-05-07T05:56:31Z
• Authors: Oriel M. M. THEKISOE; Raoul S. B. BAZIE; Andrea M. CORONEL...; Chihiro SUGIMOTO; Shin-ichiro KAWAZU; Noboru INOUE
• Keywords: 0301 basic medicine; 0303 health sciences; 03 medical and health sciences; Trypanosoma brucei brucei; Trypanosomiasis, Bovine; Animals; Cattle; DNA, Protozoan; Nucleic Acid Amplification Techniques; Sensitivity and Specificity

This study evaluated the stability of LAMP reagents when stored at 25 degrees C and 37 C, also assessed its detection efficiency on different DNA template preparations. Accordingly, using amplified in vitro cultured T. b. brucei (GUTat 3.1) from day 1 to 15 reagent storage. There were no significant differences (P>0.05) sensitivity among -20 (recommended storage temperature). above-mentioned temperatures serially diluted DNAs (genomic extracted by phenol-chloroform method, FTA card hemolysed blood) gambiense (IL2343) with high sensitivity. Reactions conducted 30 days. was poor fresh blood as added directly into reactive solution. Results this demonstrated that has potential be used field conditions for diagnosis trypanosome infections without being affected ambient tropical sub-tropical countries where trypanosomosis is endemic.