WO2004101811A2

WO2004101811A2 - Detection result storing method

Info

Publication number: WO2004101811A2
Application number: PCT/JP2004/006799
Authority: WO
Inventors: Takami Shibazaki; Toshinobu Niimura
Original assignee: Olympus Corporation
Priority date: 2003-05-16
Filing date: 2004-05-13
Publication date: 2004-11-25
Also published as: WO2004101811A3

Abstract

A detection device captures a microarray chip hybridized with a sample based on a microarray chip method using a detection unit, obtains image data digitized from an image obtained by the capture using a control unit, and outputs a result to a processing device. The processing device 3 controls the entire system using a CPU, etc., couples the image data input from the detection device, or an analytical result obtained by analyzing the image data with related information including chip information about a microarray chip input from a chip information input unit, a date and time of experiment, experimenter, sample information, etc. input from a data input device, etc., and stores then as one data file in a predetermined sample of a database of an external storage device. Thus, a database can be protected against overgrowth, and data can be easily managed and adjusted in a detection result storing method.

Description

DESCRIPTION

DETECTION RESULT STORING METHOD

Technical Field

The present invention relates to a detection result storing method for storing a detection result obtained by detecting gene using a microarray chip arranged in a matrix form on a substrate such as a glass slide, a membrane filter, etc. using a DNA, a plurality of biopolymers such as a DNA specifically coupling with protein, etc. as capture probes.

Background Art

Recently, in the field of molecular biology, an analysis relating to gene has made a remarkable progress. For example, the DNA sequences of human chromosomes are being almost completely represented. The method of analyzing gene can be determining the sequence of an unknown fragment by fixing in high density a known nucleic acid whose sequence is known as a capture probe on a substrate such as a glass slide, a membrane filter, etc., and analyzing a hybridization signal obtained by hybridizing a fragment of an unknown DNA or RNA.

In this method, using as a capture probe a known DNA which is known to be coupled with which element (sequence of gene), a sample hybridized with the capture probe can be determined to have contained what element (sequence of gene) , thereby attracting much attention as microarray chip technology.

The detection of gene in the microarray chip is normally embodied by the following processes.

PI. A sample element which is a test sample is coupled with a fluorochrome to assign a label to it. P2. The labeled^' sample is hybridized with a microarray chip in an appropriate environmental condition.

P3. The microarray chip is irradiated by the light of a wavelength exciting the fluorochrome used for a label, the amount of label material coupled with the capture probe through hybridization is photoelectrically detected, and image data is generated as the detected measurement result data. P4. The amount of irradiance of fluorescence is analyzed by detecting "the level of the intensity of the irradiance of each capture probe" from the generated image data. P5. Based on the amount of irradiance obtained by the analysis, the irradiance is compared among the capture probes and among the chips, thereby obtaining the information about a target gene. For example, between the microarray chip using a sample of a healthy person and the microarray chip using a sample of a patient suffering from cancer, the positions of irradiating capture probes are different. The capture probe (known DNA whose sequence is only known) indicating a different position relates to cancer anyhow.

In addition, since variations of types of capture probes are fixed in such a microarray chip, a plurality of elements to be measured can be detected in a single experiment.

FIG. 1A is a schematic diagram showing the image data as photoelectrically observed microarray chip processed by hybridization, and FIG. IB is a fluorescence intensity graph on the observation scanning line along A-A' shown in FIG. 1A.

As shown in FIG. 1A, the capture probes formed by 80 genes A, B, C, ... having different features are fixedly arranged in 10 rows and 8 columns as a microarray chip 1. The leftmost black-painted genes A, B, and C in the top row have the lowest brightness in the six lightness levels indicated to the left of the microarray chip 1 shown in FIG. 1A. That is, they are slightly or scarcely irradiating (few elements in the sample are coupled with these capture probes) .

The group of 10 capture probes in the observation scanning line of the third row along the line A-A' shown in FIG. IB contains the first capture probe having the highest level in the six lightness levels (highest brightness) , the second capture probe having the second lowest level in the six lightness levels, the third through fifth capture probes having the lowest level, the sixth capture probe having the second highest level, the seventh and eighth capture probes having the lowest level, the ninth capture probe having the second highest level, and the tenth capture probe having the second lowest level. That is, the sample hybridized with the microarray chip 1 contain a large number of elements^' which have coupled with at least the first, sixth, and ninth capture probes.

Normally, a practical method of producing the above-mentioned microarray chip can be fixing a solution of element-adjusted capture probes on a substrate such as a glass slide, etc. using a solution dispenser (spotter, arrayer, etc.) using a split pin and an ink jet system, and composing a sample on a substrate using a semiconductor producing technology. In either method, it is necessary to prepare the capture probe information

(which capture probe is to be fixed in which position on a chip) as design information when a microarray chip is produced.

Since a plurality of capture probes fixed on a substrate are configured to be specifically captured with specific biopolymers, it is necessary to individually identify the capture probes in the above-mentioned process P4. Therefore, in the process P4, the irradiance intensity of each capture probe can be detected according to the above-mentioned design information.

Furthermore, in the experiment using the microarray chip, only one piece of microarray chip can output a large amount of detection results, and in the molecular biology field, it is necessary to detect the status of gene for each sample in various types of samples. Therefore, it is common to prepare a database by associating the measurement result data obtained in the individual experiments conducted using the microarray chip of the same format with various types of related information such as the information, hybridization condition, analytical condition, etc. of the sample hybridized in each experiment.

Conventionally, these databases are associated with the identification information described on the label directly attached or added to the microarray chip, and the measurement result data and each piece of related information are stored in the individual databases.

It is further predicted that a quick and large capacity of genetic analysis is carried out in the microarray chip method capable of simultaneously obtaining a large number of detection results. Accordingly, it is assumed that an enormous amount of digital data of light quantity (brightness or gray scale information) and related information are required for each capture probe as image data which is the detection result information, that is, the measurement result data based on which the analysis is carried out.

Furthermore, the number of capture probes fixed in the array of a microarray chip possibly exceeds ten thousand. For example, when there are 10,000 capture probes, the chip is formed by 100 rows by 100 columns by referring to FIG. 1A. This indicates how huge the size of one microarray chip formed by the arrangement of a sample (capture probes) is.

However, since it is necessary to store the arrangement information relating to a number of fixed capture probes associated with the relationship with the obtained measurement result information in the above-mentioned microarray chip method, there is a large volume of information to be processed when the number of samples largely increases, thereby causing the overgrowth of databases.

Additionally, in an experiment and an analysis of the result of the experiment, it is not rare that the same experiment is iteratively performed to improve the precision of the detection result data of the experiments. Therefore, it is also necessary to prepare a database containing the detection result information about the experiments associated with the detection result information about the iterative experiments. Thus, the resultant database has to be managed with the correlation among plural types of data correctly maintained with an increasing amount of stored data. However, the required management is quite difficult because it requires laborious operations and a very long time.

Furthermore, for example, when a user is to communicate detection result information with another researcher, and when detection result information is to be individually moved for each microarray chip from a database, the measurement result data is to be moved together with the related information. Thus, when the measurement result data individually associated with the correlative information and stored and the related information are to be moved, the correlation according to the correlative information can be easily lost for any factor.

Thus, if the relationship between the detection result data and the related information is lost in the detection result information, and the relativeness is disconnected, the resultant measurement result data and the related information can lose the usefulness as data.

The present invention has been developed to solve the above-mentioned problems, and aims at providing a detection result storing method capable of preventing the overgrowth of a database for maintenance of the measurement result data detected in the gene detection in the microarray chip method and its related information, and easily managing and correctly arranging the data.

Disclosure of Invention

An embodiment of the detection result storing method of the present invention stores as measurement result data a detection result of a microarray chip for detection of gene, and is configured to store experiment condition information together with the measurement result data.

The measurement result data contains, for example, an image and a result of an analysis of an image .

The above-mentioned experiment condition information is configured to include at least one of, for example, arrangement information about capture probes involved in production of the microarray chip, sample information, hybridization condition information, and data analytical condition information relating to the measurement in detecting gene, and to be written to a file header portion of the measurement result data as character data. Furthermore, the above-mentioned measurement result data can be, for example, image data.

Brief Description of Drawings

FIG. 1A is a schematic diagram showing the image data obtained by photoelectrically observing a normally hybridized microarray chip; and FIG. IB is a fluorescence intensity graph in the observation scanning line A-A' shown in FIG. 1A.

FIG. 2 shows the outline of the system relating to the detection result storing method according to an embodiment of the present invention.

FIG. 3A is a flowchart for explanation of the processing operation according to the detection result storing method of the present invention used under the control of the processing device with the system configuration of the present embodiment; and

FIG. 3B shows the basic structure of the processing data.

FIG. 4 is a schematic diagram showing the data structure of the data file stored in a predetermined database storage area of an external storage device with the system configuration according to an embodiment of the present invention.

Best Mode for Carrying out the Invention FIG. 2 shows the outline of the system relating to the detection result storing method according to an embodiment of the present invention. As shown in FIG. 2, the present system comprises a detection device 2, a processing device 3, a chip information input unit 4, a data input device 5, and an external storage device 6.

The detection device 2 is gene detection apparatus in the microarray chip method, and comprises a detection unit 7 and a control unit 8. The detection unit 7 is not shown in the attached drawings, but comprises an optical system and a photoelectrical conversion device. The optical system comprises: a lens group including an objective lens, a taking lens, an intermediate focus lens, etc.; a prism group for sorting irradiating light, reflected light, an observation optical path, and a taking optical path; a mechanism group for moving a diaphragm and a lens group; a light source for generating light exciting a fluorescent material such as a laser beam source, etc. The photoelectrical conversion device comprises a CCD, a PMT, etc. The control unit 8 controls the entire detection device especially the photoelectrical conversion device of the detection unit 7, etc.

The processing device 3 comprises a CPU, etc., and controls the entire system. The processing device 3 stores the digital image data input from the detection device 2, the chip information about the microarray chip 9 input from the chip information input unit 4, the date and time of experiment, the experimenter, the sample information, etc. input from the data input device 5 in a predetermined storage area in the external storage device 6 as a data file.

The chip information input unit 4 comprises, for example, a floppy disk drive device, reads in advance chip information from the floppy disk inserted into the floppy disk drive device, manages the chip information in a memory device not shown in the attached drawings, and outputs the chip information to the processing device 3.

The chip information input unit 4 also comprises an optical reading device, etc. The optical reading device reads chip information described on the label directly attached or added to the microarray chip 9, and the read chip information is output to the processing device 3.

The data input device 5 comprises a keyboard or a data input screen, etc., and outputs the date and time of experiment, experimenter, sample information, etc. obtained in a measurement check on the microarray chip 9 externally input by the experimenter (measurement checker) as the related information relating to the measurement check.

The external storage device 6 comprises a predetermined storage area for storing a data file input from the processing device 3 as a database, and inputs/outputs a predetermined data file to and from the processing device 3 under the control of the processing device 3.

Normally, when a capture probe on the microarray chip 9 emits fluorescence in detecting gene in the above-mentioned system, it is used as the information about the amount of genes captured (coupled) by the capture probe in the used sample. To specify the gene coupled with the capture probe, the information relating to the capture probe and the sample is required. In the present embodiment, the information is incorporated into the image data of the microarray chip 9, and configured as a data file 10.

FIG. 3A is a flowchart for explanation of the processing operation based on the detection result storing method according to the present invention realized under the control of the processing device 3 with the above-mentioned system configuration, and FIG. 3B shows the basic structure of the processing data. As shown in FIG. 3A, first the detection device 2 obtains image data (SI) . In this process, the hybridized microarray chip 9 is set in the image obtaining device in the detection unit 7 of the detection device 2, and the detection unit 7 leads the exciting light from the light source to the microarray chip 9, the light from the fluorescent material of the sample coupled with each capture probe on the microarray chip 9 is leaded to the photoelectrical conversion device (for example, CCD) , the light quantity of the formed fluorescence is converted into an electrical signal by the photoelectrical conversion device, the control unit 8 generates image data 11 shown in FIG. 3B according to the converted electrical signal, and the image data 11 is obtained by the processing device 3.

The image data 11 is digital image data, and the brightness data (or gray scale data) of the digital image data relatively indicates the light quantity emitted for each capture probe which is to be checked on the microarray chip 9, that is, the amount of elements captured for each capture probe.

The microarray chip 9 set in the image obtaining device of the detection unit 7 is hybridized with the sample labeled by a fluorescent material for a predetermined time by the hybridization device not shown in the attached drawings, and then cleaned and dried under an appropriate condition as necessary. The material labeling a sample is not limited to a fluorescent material, but can be an enzyme having an irradiative substrate, a material self- irradiative by enzyme hybridization, a radioactive material, etc. The processing device 3 adds related information to the head of the obtained image data 11 (S2) . In this process, as shown in FIG. 3B, the chip information about the microarray chip 9 input from the chip information input unit 4 and the test information input from the data input device 5 are added to the head of the image data 11 as related information data 12 relating to the above-mentioned gene detecting process.

The chip information is, for example, the information about the DNA sequence, etc. fixed on the substrate of the microarray chip 9, and the test information is, for example, the date and time of experiment, experimenter, sample information, hybridization condition, taking condition, etc., which is character information containing symbols and numerals.

The processing device 3 finally stores the data file with the related information data 12 added to the head of the image data 11 in the storage device (S3) . In this process, the entire data with the related information data 12 added to the head of the image data 11 is stored as one electronic data file. Therefore, file information is further added, and the entire data is stored as one data file relating to the detection of gene on the microarray chip 9 in a predetermined storage area of the database of the external storage device 6 shown in FIG. 2.

Thus, a detection result formed by the image data 11 relating to the detection of gene on the microarray chip 9 and the related information data 12 is stored as one electronic data file in the database.

FIG. 4 is a schematic diagram showing the data structure of the data file 10 shown in FIG. 2 and stored in a storage area of the database of the external storage device 6. As shown in FIG. 4, the data file 10 is configured by adding file information 13 to the head of the detection result data comprising the image data 11 and the related information data 12 shown in FIG. 3B. The file information 13 is, for example, identification information designating the microarray chip 9, etc.

When the above-mentioned data file 10 is searched for and extracted from the database, not only the file information 13, but also the date and time of experiment, experimenter, sample information, hybridization condition, signal detection condition, etc. can be used as a retrieval key to search for and extract a data file.

Since the image data 11, the related information data 12, and the file information 13 are incorporated into one data file, the data can be arbitrarily moved and copied without disconnecting the image data 11, the related information data 12, and the file information 13, or without a possible disadvantage, thereby constantly utilizing the useful gene detection data. According to the present invention, the character data can be incorporated as is into image data 11 as the related information data 12, but each piece of data can also be coupled after being treated in a common reversible compression process. In addition, according to the present invention, the related information data 12 is added before the image data 11, but it is obvious that the related information data 12 can also be added after the image data 11.

However, when it is assumed that the data structure of the related information data 12 to be added to the image data 11 is changed, it is preferable to write the related information data 12 to the head of the file as character data as shown in FIG. 3B to easily change the data structure. Furthermore, although the experiment information contains the date and time of experiment, experimenter, sample information, hybridization condition, and taking condition as the related information data 12, the sample condition is, for example, the information about the derivation of a sample, and refers to when, how (in which state, healthy, ill, or what diseases) , and who takes the sample.

A hybridization condition refers to, for example, the temperature of a sample, a stirring condition of a sample, for example, a solution drive speed, an amount of solution drive, a hybridizing time, the solvent condition of pH and sodium density, etc. A taking condition refers to, for example, an arrangement condition and an observation condition. An arrangement condition includes the position data (for example, the coordinates data of each capture probe relative to the reference position) of a capture probe, that is, the arrangement data of the capture probe. An observation condition refers to, for example, an exposing time, an observation wavelength, exciting light intensity, a exciting wavelength, etc.

In addition to the above-mentioned data, a data analytical condition can also be included. A data analytical condition can be, for example, the reference data for use in normalizing the fluorescence intensity as digital data as shown in the bar graph data shown in FIG. IB, and the method of calculating the fluorescence intensity, etc. According to the present embodiment, image data is used as measurement result data in the explanation above. However, it is not limited to image data, but the present invention can be applied by using a result of analyzing image data as measurement result data.

As described above, according to the present invention, the image data which is detection result data obtain in gene detecting process is stored together with the related information as one file. Therefore, the database for maintenance of the measurement result and the related information obtained in detecting gene in the microarray chip method can be protected against overgrowth, and the data can be easily managed and adjusted.

Claims

1. A detection result storing method for storing a detection result of a microarray chip in detecting gene as measurement result data together with experiment condition information.

2. The detection result storing method according to claim 1, wherein said experiment condition information comprise at least one of arrangement information about capture probes, sample information, hybridization condition information, data analytical condition information relating to measurement in detecting gene when the microarray chip is generated.

3. The detection result storing method according to claim 1, wherein said experiment condition information is written as character to a file header portion of the measurement result data.

4. The method according to claim 1, wherein said measurement result data is image data.