Loop-Mediated Isothermal Amplification Test for Trypanosoma gambiense Group 1 with Stem Primers: A Molecular Xenomonitoring Test for Sleeping Sickness

Figure 2

(a) The tube scanner fluorometric output of stem LAMP (red) and standard LAMP (blue) tests using ~10 pg of DNA from isolate PT41 () and DNA prepared from CSF () of a confirmed HAT patient. The unit reports the fluorescence in millivolts (mV) on -axis and time in minutes on the -axis. (b) The acquisition of melts curves for PT41, DNA prepared from CSF, and supernatant prepared from a tsetse fly sample spiked with B014 DNA. was 89.5°C for PT41 and DNA from CSF patient and 89.4°C for B014 in tsetse fly sample. The nonspecific product (FP) is induced for illustration and showed that ranged from 72 to 76°C and 90 to 92°C (c). The duplication of melts peaks for PT41, CSF, and B014 DNA samples using Rotorgene 6000. The melt peaks showed of ~89°C for PT41 and CSF DNA and ~ 88°C for B014. (d) The amplicons for stem LAMP test (I) without outer primers and (II) with outer primers.

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