JP2008035773A5 - - Google Patents

Claims (13)

  Detection of mites or molds in which a nucleic acid probe capable of hybridizing to a detection target region amplified using mite or mold-derived genomic DNA or RNA collected from a test target as a template is immobilized on a carrier Or an identification microarray.   Nucleic acid probes are derived from Dermatophagoides farinae, Dermatophagoides pteronyssinus or Tyrophagus pterescentiae as the mites, and Alternaria alternata, Aspergills restricts, Aspergills restrictus, Aspergills The microarray according to claim 1, which can hybridize to a detection target region amplified using genomic DNA or RNA derived from Aspergillus nigar, Penicillium chrysogenum, Fusarium sp. Or Wallemia sebi as a template.   The microarray according to claim 1 or 2, wherein the detection target region is amplified using a tick gene-specific primer set or a mold gene-specific primer set.   The microarray according to any one of claims 1 to 3, wherein the tick gene-specific primer set or the mold gene-specific primer set has a continuous 15 to 30 base sequence. The tick gene-specific primer set is the following (a), (b) or (c):
(A) A set of a primer consisting of the base sequence shown in SEQ ID NO: 56 and a primer consisting of the base sequence shown in SEQ ID NO: 57 (b) Substituting, deleting or deleting 1 to 5 bases in the base sequence shown in SEQ ID NO: 56 A primer consisting of an added base sequence and a primer consisting of a base sequence in which 1 to 5 bases are substituted, deleted or added in the base sequence of SEQ ID NO: 57, and the same region as the primer set of (a) above (C) a primer consisting of a nucleic acid fragment capable of hybridizing under stringent conditions to a nucleic acid fragment consisting of a base sequence complementary to the base sequence shown in SEQ ID NO: 56, and SEQ ID NO: 57 Nuclei capable of hybridizing under stringent conditions to a nucleic acid fragment consisting of a base sequence complementary to the indicated base sequence Consists of a primer consisting of a fragment, the a primer set capable of amplifying the same area as the primer set (a), according to claim 3 microarray according. The mold gene-specific primer set is the following (a), (b) or (c):
(A) A set of a primer consisting of the base sequence shown in SEQ ID NO: 58 and a primer consisting of the base sequence shown in SEQ ID NO: 59 (b) Substituting, deleting or deleting 1 to 5 bases in the base sequence shown in SEQ ID NO: 58 A region consisting of a primer consisting of an added base sequence and a primer consisting of a base sequence obtained by substituting, deleting or adding 1 to 5 bases in SEQ ID NO: 59 base sequence, the same region as the primer set of (a) above (C) a primer consisting of a nucleic acid fragment capable of hybridizing under stringent conditions to a nucleic acid fragment consisting of a base sequence complementary to the base sequence shown in SEQ ID NO: 58 and SEQ ID NO: 59 Nucleic acid capable of hybridizing under stringent conditions to a nucleic acid fragment consisting of a base sequence complementary to the indicated base sequence 4. The microarray according to claim 3, wherein the microarray is a set of primers that are composed of fragments and that can amplify the same region as the primer set of (a).   The nucleic acid probe comprises at least one base sequence selected from the group consisting of SEQ ID NOs: 1, 5, 9, 13, 17 and 21, or a part or all of a base sequence complementary to the base sequence. 2. The microarray according to 2.   The nucleic acid probe is at least one base sequence selected from the group consisting of SEQ ID NOs: 24, 28, 32, 36, 40, 44, 48-53 and 114-121, or a part of a base sequence complementary to the base sequence, or The microarray according to claim 1 or 2, comprising all of them.   The microarray according to claim 7 or 8, wherein at least one base sequence or a part of a base sequence complementary to the base sequence is a continuous 10 to 30 base sequence.   The microarray according to any one of claims 1 to 9, wherein the carrier has a carbon layer and a chemical modification group on a surface thereof.   The microarray according to claim 10, wherein the carbon layer is a diamond-like carbon layer, and the chemically modifying group is an active ester group. Collecting dust from the inspection object;
Separating mites and molds from the collected dust, and extracting genomic DNA or RNA from the separated mites and molds;
Using the extracted genomic DNA or RNA as a template, and amplifying the detection target region;
A method for detecting or identifying mites or molds, comprising a step of detecting a detection target region using the microarray according to any one of claims 1 to 11. The at least one selected from the nucleic acid probe which consists of a base sequence in any one of sequence number 62-93, 98-113, 122-153, 158-205, 210-225 is fix | immobilized by the support | carrier. The microarray according to any one of 6, 10, and 11.