Pcr diagnostics of dermatophytes and other pathogenic fungi

A method of detecting fungi infections in patient samples comprising the following steps:

• (A) extracting nucleic acid from the sample by:
• (a) heating a sample containing the fungi in a lysis buffer to between 80 and 100° C. for 10 minutes;
• and
• (b) mixing the solution with a neutralizing buffer;
• wherein said lysis buffer consists of, in aqueous solution:
• (i) a reducer selected from the group consisting of sodium carbonate, sodium sulfate, beta-mercaptoethanol, dithiothreitol, sodium sulfide, sodium chloride, sodium chlorite, sodium iodate, and sodium bicarbonate;
• (ii) a salt selected from the group consisting of potassium chloride and ammonium persulfate;
• and (iii) a buffering compound selected from the group consisting of CAPSO, TRIS, HEPES, and phosphate buffer;
• wherein said neutralizing buffer is an aqueous 0.5-3% w/v protein solution;
• (B) amplifying the nucleic acid with PCR using fungus-specific primers;
• and
• (C) detecting fungus-specific genes.

A multiplex PCR kit for extracting nucleic acid from fungi and detecting dermatophytes comprising:

• (a) a lysis buffer consisting of, in aqueous solution:
• (i) a reducer selected from the group consisting of sodium carbonate, sodium sulfate, beta-mercaptoethanol, dithiothreitol, sodium sulfide, sodium chloride, sodium chlorite, sodium iodate, and sodium bicarbonate;
• (ii) a salt selected from the group consisting of potassium chloride and ammonium persulfate;
• and (iii) a buffering compound selected from the group consisting of CAPSO, TRIS, HEPES, and phosphate buffer;
• (b) a neutralizing buffer, wherein said neutralizing buffer is an aqueous 0.5-3% w/v protein solution;
• (c) fungus-specific primers selected from the group consisting of panDerm1 (SEQ ID NO:
• 1), panDerm2 (SEQ ID NO:
• 2), uni (SEQ ID NO:
• 3), Trubrum-rev (SEQ ID NO:
• 4), Ef-rev (SEQ ID NO:
• 5), Micr773-for (SEQ ID NO:
• 10), Micr885-rev (SEQ ID NO:
• 11), Trichopyros-for (SEQ ID NO:
• 12), and Trichopyros-rev (SEQ ID NO:
• 13);
• and
• (d) agents for performing a PCR reaction.