Development of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Trichosporon asahii in Experimental and Clinical Samples

Figure 3

Sensitivity of LAMP method using serially diluted genomic DNA (1, 3.56 * 10⁶ copies, 2, 3.56 * 10⁵ copies, 3, 3.56 * 10⁴ copies, 4, 3.56 * 10³ copies, 5, 3.56 * 10² copies, 6, 3.56 * 10¹ copies, 7, 3.56 copies) with T. asahii CBS2479 as template. The detection limit for the assay was 3.56 * 10³ genomic copies. (a) Sensitivity of the LAMP assay was monitored by real-time measurement of turbidity. (b) Sensitivity of LAMP for T. asahii detection was visualized by gel electrophoresis. The positive reaction was seen as a ladder-like pattern on 1% agarose gel electrophoresis analysis. (c) Sensitivity of PCR for T. asahii detection was visualized by gel electrophoresis. The positive reaction was seen as a ladder-like pattern on 1% agarose gel electrophoresis analysis. Marker, DL2000 DNA marker. (d) The specificity of LAMP for positive detection by naked eye detection. Green colour was observed using the naked eye in the tube which contained T. asahii, whereas others remained light orange after the reaction.

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