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

Figure 2

Specificity of LAMP detection for T. asahii. (a), (b) Lane 1: negative control; lane 2: T. asahii isolate CBS2479; lane 3: Trichosporon domesticum; lane 4: Trichosporon dermatis; lane 5: Trichosporon jirovecii; lane 6: Trichosporon mucoides; lane 7: Trichosporon asteroides; lane 8: Trichosporon coremiiforme; lane 9: Trichosporon inkin; lane 10: Trichosporon japonicum; lane 11: Trichosporon lactis; lane 12: Trichosporon ovoides; lane 13: Trichosporon dohaense; lane 14: Trichosporon faecale; lane 15: Trichosporon debeurumanianum; lane 16: Trichosporon montevideense. (a) Specificity of the LAMP assay monitored by real-time measurement of turbidity (LA-320C). Positive reaction was observed in T. asahii, whereas none of other Trichosporon spp. showed turbidity increases. (b) The specificity of LAMP for T. asahii 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. (c), (d) Lane 1: negative control, lane 2: T. asahii isolate CBS2479; lane 3: Cryptococcus gattii; lane 4: Cryptococcus laurentii; lane 5: Cryptococcus luteolus; lane 6: Cryptococcus neoformans var. grubii; lane 7: Cryptococcus neoformans var. neoformans; lane 8: Cryptococcus podzolicus; lane 9: Cryptococcus podzolicus; lane 10: Pichia pastoris; lane 11: Candida albicans; lane 12: Candida glabrata; lane 13: Candida krusei; lane 14: Candida parapsilosis; lane 15: Candida parapsilosis; lane 16: Candida stellata; lane 17: Candida tropicalis; lane 18: Debaryomyces hansenii; lane 19: Fonsecaea pedrosoi; lane 20: Exophiala dermatitidis; lane 21: Filobasidiella neoformans var. neoformans; lane 22: Sporothrix schenckii; lane 23: Acremonium chrysogenum; lane 24: Aspergillus fumigatus; lane 25: Aureobasidium pullulans. (c) Specificity of the LAMP assay monitored by real-time measurement of turbidity (LA-320C). Positive reaction was observed in T. asahii, whereas none of other non-Trichosporon fungal strains showed turbidity increases. (d) The specificity of LAMP for T. asahii detection by naked eye detection. Light blue 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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