Application of laser capture microdissection and polymerase chain reaction in the diagnosis of Trichoderma longibrachiatum infection: a promising diagnostic tool for 'fungal contaminants' infection.

ABSTRACT

Although Trichoderma species are usually considered to be culture contaminants, an increasing number of case reports have demonstrated their pathogenicity. Current diagnostic tools, including fungal culture, radiology, histopathology, and direct microscopy examination, are often unable to differentiate the pathogenicity of 'fungal contaminants' such as Trichoderma species in patients. Accurate diagnostic tools for 'fungal contaminants' infection have become the urgent needs. To that end, we applicated laser capture microdissection (LCM) and polymerase chain reaction (PCR) to confirm T. longibrachiatum infection for the first time. A 57-year-old man presented with a cough and hemoptysis lasting for more than 40 days. Computed tomography scan revealed a mass at the left hilum. In addition to pulmonary spindle cell carcinoma, fungal hyphae were also detected in histopathological examination. The cultured fungus was identified as T. longibrachiatum using molecular procedures. The results from DNA sequencing of DNA obtained by LCM revealed the identical result. Antifungal susceptibility testing revealed resistance to itraconazole, fluconazole and flucytosine. The patient was managed with oral voriconazole for 4 months. No relapse of Trichoderma infection was observed at a year follow-up visit. Although there are potential disadvantages, LCM-based molecular biology technology is a promising diagnostic tool for 'fungal contaminants' infection.