Detecting method and detection substrate for use therein

#The present invention is directed to examining multiple specimens at a time for multiple items, and provides a method in which matrix substrates with biological samples having different properties and origins bound thereto are prepared, and on each matrix region, oligonucleotides having different sequences, proteins or drugs are spotted in an array, whereby multiple specimens are examined at a time for multiple items.

#The present invention also relates to a method in which by using an oligonucleotide having a known base sequence as a detection probe to detect whether a complex is formed by intermolecular bond with this oligonucleotide, detection is made whether or not components having a capability of bonding to the above described detection probe are contained, and a detection substrate having the oligonucleotide as a detection probe fixed on its surface, which is used exclusively for this detecting method.

#2. Related Background Art

#FIG. 1 shows one example of an arrangement aspect of defined regions on a substrate in the present invention;

#FIGS. 2A and 2B show one example of matrices in the present invention, wherein FIG. 2A is a plan view, and FIG. 2B is a 2B-2B sectional view thereof;

#FIG. 3 is a schematic explanatory view of a specimen solution discharging method by bubble jet process that is an embodiment of the present invention;

#One embodiment of the present invention will be described below referring to FIG. 1. FIG. 1 shows a substrate surface with 64 defined regions formed thereon, wherein each region (matrix) measures 1 mm by 1 mm, and a space x between regions can be selected freely. For methods of preparing biological sample binding matrix substrates, for example, a method can be used in which the solution of a first sample (e.g. biological sample) is printed on the entire surface of defined regions on the substrate as a “solid print pattern” by coating and ink jet processes, or is supplied by methods such as chemical synthesis on the substrate, and is bound in a matrix form on the substrate through adsorption to the substrate or chemical reaction between functional groups existing in the biological sample and functional groups existing on the substrate. Furthermore, the situation in which the first sample is bound on the entire surface of defined regions means a situation in which the first sample is bound across the entire surface such that when a second sample and samples thereafter are supplied in these defined regions, these reactions occur without being limited to the positions in the above described regions in which the samples are supplied. For example, the first sample may be fixed in layered form on the entire surface, or the masses of molecules constituting the first sample may be dispersed on the entire surface in high density with micro-spaces being kept among them.

#The defined regions on this substrate may previously be provided on the substrate as a well constituted by sections separated in pattern form by walls of hydrophobic compounds.

#Also, when using a substrate with nucleoside acid (cDNA) being a biological sample fixed thereon as the first sample, two or more probe DNAs possibly included in cDNA are contacted with cDNA on the substrate as the second sample and samples thereafter, and products of reaction with the above described probes are detected on the above described solid phase to detect the presence or absence of probe DNA sequences in the above described cDNA, two or more probes are supplied in an array form as mutually independent spots in each matrix with various kinds of CDNA bound in the defined regions, thereby making it possible to perform simultaneous detection with two or more probes.