Method for the simultaneous detection and differentiation of the following different fungal pathogenic species Candida albicans, Candida parapsilosis, Candida tropicalis, Candida krusei, Candida glabrata, Aspergillus flavus, Aspergillus versicolor, Aspergillus nidulans, Aspergillus fumigatus, and Cryptococcus neoformans in combination with each other, in one single assay, including; (i) releasing, isolating and/or concentrating the nucleic acids of the fungal pathogens possibly present in the sample, (ii) if necessary, amplifying the Internal Transcribed Spacer region of said nucleic acids with at least one fungal universal primer pair, (iii) hybridizing the nucleic acids of step (i) or (ii) with the following ITS-1 species specific oligonucleotide probes, these probes being selected to detect and differentiate said different fungal pathogenic species, comprising;_ - for Candida albicans: at least one of GTCTAAACTTACAACCAATT (SEQ ID NO 1), TGTCACACCAGATTATTACT (SEQ ID NO 2), or TATCAACTTGTCACACCAGA (SEQ ID NO 3), - for Candida parapsilosis: TGCCAGAGATTAAACTCAAC (SEQ ID NO 5), - for Candida tropicalis: GGTTATAACTAAACCAAACT (SEQ ID NO 6), - for Candida krusei: GGAATATAGCATATAGTCGA (SEQ ID NO 9), - for Candida glabrata: GAGCTCGGAGAGAGACATC (SEQ ID NO 10), - for Cryptococcus neoformans: CATCTACACCTGTGAACTGT (SEQ ID NO 15), - for Aspergillus flavus: GTTTACTGTACCTTAGTTGCT (SEQ ID NO 18), - for Aspergillus versicolor: CCTCTCGGGGGCGAGCC (SEQ ID NO 21), - for Aspergillus nidulans: CCGAGTGCGGCTGCCTC (SEQ ID NO 22), - for Aspergillus fumigatus: GTTGATTATCGTAATCAGT (SEQ ID NO 26), or variants of said probes, said variants differing from the sequences cited above by the deletion and/or addition of one or two nucleotides at the 5' and/or 3' extremity of the nucleotide sequence, without affecting the species specific hybridization behavior of the probe, or the RNA equivalents of said probes, wherein T is replaced by U, or the complementary molecules of said probes, and (iv) detecting the hybridization complexes formed in step (iii). (i) releasing, isolating and/or concentrating the nucleic acids of the fungal pathogens possibly present in the sample, (ii) if necessary, amplifying the Internal Transcribed Spacer region of said nucleic acids with at least one fungal universal primer pair, (iii) hybridizing the nucleic acids of step (i) or (ii) with the following ITS-1 species specific oligonucleotide probes, these probes being selected to detect and differentiate said different fungal pathogenic species, comprising;_ - for Candida albicans: at least one of GTCTAAACTTACAACCAATT (SEQ ID NO 1), TGTCACACCAGATTATTACT (SEQ ID NO 2), or TATCAACTTGTCACACCAGA (SEQ ID NO 3), - for Candida parapsilosis: TGCCAGAGATTAAACTCAAC (SEQ ID NO 5), - for Candida tropicalis: GGTTATAACTAAACCAAACT (SEQ ID NO 6), - for Candida krusei: GGAATATAGCATATAGTCGA (SEQ ID NO 9), - for Candida glabrata: GAGCTCGGAGAGAGACATC (SEQ ID NO 10), - for Cryptococcus neoformans: CATCTACACCTGTGAACTGT (SEQ ID NO 15), - for Aspergillus flavus: GTTTACTGTACCTTAGTTGCT (SEQ ID NO 18), - for Aspergillus versicolor: CCTCTCGGGGGCGAGCC (SEQ ID NO 21), - for Aspergillus nidulans: CCGAGTGCGGCTGCCTC (SEQ ID NO 22), - for Aspergillus fumigatus: GTTGATTATCGTAATCAGT (SEQ ID NO 26), - for Candida albicans: at least one of GTCTAAACTTACAACCAATT (SEQ ID NO 1), TGTCACACCAGATTATTACT (SEQ ID NO 2), or TATCAACTTGTCACACCAGA (SEQ ID NO 3), at least one of GTCTAAACTTACAACCAATT (SEQ ID NO 1), TGTCACACCAGATTATTACT (SEQ ID NO 2), or TATCAACTTGTCACACCAGA (SEQ ID NO 3), - for Candida parapsilosis: TGCCAGAGATTAAACTCAAC (SEQ ID NO 5), TGCCAGAGATTAAACTCAAC (SEQ ID NO 5), - for Candida tropicalis: GGTTATAACTAAACCAAACT (SEQ ID NO 6), GGTTATAACTAAACCAAACT (SEQ ID NO 6), - for Candida krusei: GGAATATAGCATATAGTCGA (SEQ ID NO 9), GGAATATAGCATATAGTCGA (SEQ ID NO 9), - for Candida glabrata: GAGCTCGGAGAGAGACATC (SEQ ID NO 10), GAGCTCGGAGAGAGACATC (SEQ ID NO 10), - for Cryptococcus neoformans: CATCTACACCTGTGAACTGT (SEQ ID NO 15), CATCTACACCTGTGAACTGT (SEQ ID NO 15), - for Aspergillus flavus: GTTTACTGTACCTTAGTTGCT (SEQ ID NO 18), GTTTACTGTACCTTAGTTGCT (SEQ ID NO 18), - for Aspergillus versicolor: CCTCTCGGGGGCGAGCC (SEQ ID NO 21), CCTCTCGGGGGCGAGCC (SEQ ID NO 21), - for Aspergillus nidulans: CCGAGTGCGGCTGCCTC (SEQ ID NO 22), CCGAGTGCGGCTGCCTC (SEQ ID NO 22), - for Aspergillus fumigatus: GTTGATTATCGTAATCAGT (SEQ ID NO 26), GTTGATTATCGTAATCAGT (SEQ ID NO 26),