Patent ID: 7906286
Filing Date: 2011-03-15
Classification: C12Q,G01N,Y10S

Abstract:
1. A method for identifying a fungal species, comprising: extracting a nucleic acid from an affected tissue; amplifying the extracted nucleic acid using a primer set capable of amplifying at least a portion of an ITS region of fungi; and obtaining information about a partial sequence of the amplified nucleic acid which permits fungal species identification and determining and identifying a fungal species of the amplified nucleic acid, wherein the fungal species of the amplified nucleic acid is determined and identified by using a probe set comprising a plurality of nucleic acid probes, at least one of which is designed from a partial sequence specific to the fungal species of the amplified nucleic acid, wherein a hybridization reaction is performed with the at least one of the plurality of nucleic acid probes, and the fungal species of the amplified nucleic acid is determined and identified from a signal intensity obtained from the hybridization reaction, and wherein the plurality of nucleic acid probes comprises: (1) a first probe belonging to any group selected from the following groups 1 to 29 and a second probe belonging to any group selected from the following groups 1 to 29 and not belonging to the group to which the first probe belongs, or (2) a third probe consisting of the complementary sequence of the first probe and a fourth probe consisting of the complementary sequence of the second probe: group 1: (1) a probe consisting of tctttgaaacaaacttgctttggcgg (SEQ ID NO: 1), (2) a probe consisting of ccgccagaggtctaaacttacaacc (SEQ ID NO: 2), (3) a probe consisting of gacggtagtggtaaggcgggat (SEQ ID NO: 3), and (4) a probe consisting of ggcggtaacgtccaccacgtat (SEQ ID NO: 4), group 2: (1) a probe consisting of tgtgttttgttctggacaaacttgctttg (SEQ ID NO: 5), (2) a probe consisting of ctgccgccagaggacataaacttac (SEQ ID NO: 6), (3) a probe consisting of tagtggtataaggcggagatgcttga (SEQ ID NO: 7), and (4) a probe consisting of tctggcgtcgcccattttattcttc (SEQ ID NO: 8), group 3: (1) a probe consisting of ggtgttttatcacacgactcgacact (SEQ ID NO: 9), (2) a probe consisting of ggagttctcccagtggatgcaaac (SEQ ID NO: 10), (3) a probe consisting of ggccatatcagtatgtgggacacg (SEQ ID NO: 11), and (4) a probe consisting of aggttttaccaactcggtgttgatctag (SEQ ID NO: 12), group 4: (1) a probe consisting of gcttaactgcgcggcgaaaaac (SEQ ID NO: 13), (2) a probe consisting of agataggttgggccagaggtttaaca (SEQ ID NO: 14), (3) a probe consisting of tcttagtcggactaggcgtttgctt (SEQ ID NO: 15), and (4) probe consisting of tcgttgaatggtgtggcggat (SEQ ID NO: 16), group 5: (1) a probe consisting of gtgttgccttccgaaatatcacagttg (SEQ ID NO: 17), and (2) a probe consisting of cagttgtcgcaatacgttacttcaacttt (SEQ ID NO: 18), group 6: (1) a probe consisting of gcggccagttcttgattctctgc (SEQ ID NO: 19), (2) a probe consisting of agctcgtctctccagtggacataaac (SEQ ID NO: 20), (3) a probe consisting of ttgaaagtggctagccgttgcc (SEQ ID NO: 21), and (4) consisting of tcgtggtaagcttgggtcatagagac (SEQ ID NO: 22), group 7: (1) a probe consisting of agcggaacgaaaacaacaacacct (SEQ ID NO: 23), (2) a probe consisting of acctagtgtgaattgcagccatcg (SEQ ID NO: 24), (3) a probe consisting of gacgtgtaaagagcgtcggagc (SEQ ID NO: 25), and (4) a probe consisting of gcgagtgttgcgagacaacaaaaag (SEQ ID NO: 26), group 8: (2) a probe consisting of aggcgttgctccgaaatatcaacc (SEQ ID NO: 28), group 9: (1) a probe consisting of tggggcctgccagagattaaact (SEQ ID NO: 29), (2) a probe consisting of gtgttgagcgatacgctgggttt (SEQ ID NO: 30), and (3) a probe consisting of gttttttccactcattggtacaaactcca (SEQ ID NO: 31), group 10: (1) a probe consisting of accgccagaggttataactaaaccaaa (SEQ ID NO: 32), (2) a probe consisting of gagcaatacgctaggtttgtttgaaagaa (SEQ ID NO: 33), and (3) a probe consisting of acgcttattttgctagtggccacc (SEQ ID NO: 34), group 11: (1) a probe consisting of tgaactgttgattgacttcggtcaattga (SEQ ID NO: 35), and (2) a probe consisting of gcgtgtttaacttgtcttatctggcg (SEQ ID NO: 36), group 12: (1) a probe consisting of gttctactacttgacgcaagtcgagt (SEQ ID NO: 37), and (2) a probe consisting of ttgggcgtctgcgatttctgatc (SEQ ID NO: 38), group 13: (1) a probe consisting of caacggatctcttggcttccaca (SEQ ID NO: 39), and (2) a probe consisting of ttgagagtcatgaaaatctcaatccctcg (SEQ ID NO: 40), group 14: (1) a probe consisting of cccgtgtctatcgtaccttgttgc (SEQ ID NO: 41), (2) a probe consisting of tgaacgctgttctgaaagtatgcagt (SEQ ID NO: 42), and (3) a probe consisting of gccagccgacacccaactttatt (SEQ ID NO: 43), group 15: (1) a probe consisting of cccatccgtgtctattgtaccctgt (SEQ ID NO: 44), (2) a probe consisting of acacgaacactgtctgaaagcgtg (SEQ ID NO: 45), and (3) a probe consisting of cctgccgacgttttccaaccat (SEQ ID NO: 46), group 16: (1) a probe consisting of tctctctgaatgctggacggtgtc (SEQ ID NO: 47), (2) a probe consisting of ctcgccgaaggagtgattctcaga (SEQ ID NO: 48), (3) a probe consisting of ttccaccgggagaggagaaagg (SEQ ID NO: 49), and (4) a probe consisting of acaaaaccagcgccttcaggac (SEQ ID NO: 50), group 17: (2) a probe consisting of cgccggaggattactctggaaaac (SEQ ID NO: 52), group 18: (1) a probe consisting of gtccggggacaatcaactccct (SEQ ID NO: 53), (2) a probe consisting of aatccatgaatactgttccgtctgage (SEQ ID NO: 54), and (3) a probe consisting of ggccggttttctggcctagtttt (SEQ ID NO: 55), group 19: (1) a probe consisting of agcctctttgggggctttagct (SEQ ID NO: 56), (2) a probe consisting of acagacatcaaaaaatcttggaaagctgt (SEQ ID NO: 57), (3) a probe consisting of ctgggcgaatgggcagtcaaac (SEQ ID NO: 58), and (4) a probe consisting of ctctggccttcccccaaatctc (SEQ ID NO: 59), group 20: (1) a probe consisting of agacaccaagaaaaattctctgaagage (SEQ ID NO: 60), (2) a probe consisting of gaatgggcagccaattcagcgc (SEQ ID NO: 61), and (3) a probe consisting of cttctgggagcctcgagccg (SEQ ID NO: 62), group 21: (1) a probe consisting of cggcgagcctctctttatagcg (SEQ ID NO: 63), (2) a probe consisting of cctctctttatagcggctcaacgc (SEQ ID NO: 64), and (3) a probe consisting of ggctttctaggcgaatgggcaa (SEQ ID NO: 65), group 22: (1) a probe consisting of aggacagacatcaaaaaatcttgaagagc (SEQ ID NO: 66), and (2) a probe consisting of aagctcggcttgtgtgatggac (SEQ ID NO: 67), group 23: (1) a probe consisting of acaccaaggaaaattctctgaagggc (SEQ ID NO: 68), and (2) a probe consisting of ccaaggaaaattctctgaagggctgt (SEQ ID NO: 69), group 24: (1) a probe consisting of tctctttagtggctcaacgctgga (SEQ ID NO: 70), and (2) a probe consisting of ggacagacgcaaaaaaattctttcagaag (SEQ ID NO: 71), group 25: (1) a probe consisting of tgggcaataaccagcgcctcta (SEQ ID NO: 72), and (2) a probe consisting of tcagggatgcatttctctgcgaatc (SEQ ID NO: 73), group 26: (1) a probe consisting of cctctctttagtggctaaacgctgg (SEQ ID NO: 74), and (2) a probe consisting of cgccctggcctcaaaatctgtt (SEQ ID NO: 75), group 27: (1) a probe consisting of ttcgagcgtcatttcaacccctc (SEQ ID NO: 76), group 28: (1) a probe consisting of gttgacctcggatcaggtagggat (SEQ ID NO: 77), and group 29: (1) a probe consisting of aactttcaacaacggatctcttggttct (SEQ ID NO: 78), (2) a probe consisting of gcatcgatgaagaacgcagcga (SEQ ID NO: 79)), and (3) a probe consisting of gtgaatcatcgaatctttgaacgcaca (SEQ ID NO: 80).