JP2014007995A5

JP2014007995A5 -

Info

Publication number: JP2014007995A5
Application number: JP2012146470A
Authority: JP
Filing date: 2012-06-29
Publication date: 2015-07-09
Anticipated expiration: 2032-06-29

Claims

A method for comparatively analyzing the expression level of small RNA between multiple samples,
A measurement step of measuring the expression levels of a plurality of mRNAs simultaneously with the measurement of the expression levels of a plurality of small RNAs for each of the plurality of specimens;
A representative value acquisition step of acquiring a representative value represented by a logarithmic value from the measured value of the mRNA expression level for each specimen;
The arbitrarily selected first sample is used as a reference sample, and the difference between the mRNA representative value of the reference sample and the mRNA representative values of the remaining second and subsequent samples is used as a correction coefficient for each of the second and subsequent samples. Acquiring a correction coefficient;
Correcting the small RNA expression level measured in the second and subsequent samples using the obtained correction coefficients for the second and subsequent samples, respectively; and a small RNA expression level in the reference sample and the second Including a comparison step that compares the corrected small RNA expression level in subsequent samples,
In the correction coefficient acquisition step, when a value obtained by subtracting the mRNA representative value of the second and subsequent samples from the mRNA representative value of the reference sample is acquired as a correction coefficient, the small RNA expression level in the second and subsequent samples is acquired in the correction step. The correction is performed by adding a correction coefficient to the logarithmic value of the measured value,
In the correction coefficient acquisition step, when a value obtained by subtracting the mRNA representative value of the reference sample from the mRNA representative value of the second and subsequent samples is acquired as a correction coefficient, the small RNA expression level in the second and subsequent samples is acquired in the correction step. The method, wherein the correction is performed by subtracting a correction coefficient from the logarithmic value of the measured value.

The method according to claim 1, wherein the expression level of the small RNA is comparatively analyzed between the two specimens.

In the measurement step, a plurality of small RNA capture probes and a plurality of mRNA capture probes immobilized on a support are brought into contact with a nucleic acid sample derived from a sample labeled with a labeling substance, and hybridization is performed. And measuring the expression level of each small RNA and each mRNA as a signal intensity measurement value by measuring a signal from a labeling substance in each probe-immobilized region on the support, respectively. The method described.

The method according to claim 3, wherein the probe is a nucleic acid probe and the labeling substance is a fluorescent dye.

The method according to any one of claims 1 to 4, wherein the representative value is an average value calculated by a logarithmic value of the mRNA expression level measurement value.

The method according to any one of claims 1 to 5, wherein the small RNA is miRNA.

A small RNA expression analyzer for comparative analysis of the expression level of small RNA in multiple specimens,
For each of a plurality of specimens, storage means for storing a plurality of small RNA expression levels and a plurality of mRNA expression levels measured simultaneously,
For each sample, representative value acquisition means for acquiring a representative value represented by a logarithmic value from the measured value of the mRNA expression level,
The arbitrarily selected first sample is used as a reference sample, and the difference between the mRNA representative value of the reference sample and the mRNA representative values of the remaining second and subsequent samples is used as a correction coefficient for each of the second and subsequent samples. Correction coefficient acquisition means for acquiring,
Correction means for correcting the small RNA expression level measured in the second and subsequent samples, respectively, using the correction coefficients for the second and subsequent samples acquired by the correction coefficient acquiring means;
An output means for outputting a comparison result between the small RNA expression level of the reference sample and the corrected small RNA expression level in the second and subsequent samples,
When the correction coefficient acquisition unit acquires a value obtained by subtracting the mRNA representative value of the second and subsequent samples from the mRNA representative value of the reference sample as the correction coefficient, the correction unit obtains the small RNA expression level in the second and subsequent samples. The correction is performed by adding a correction coefficient to the logarithmic value of the measured value,
When the correction coefficient acquisition unit acquires a value obtained by subtracting the mRNA representative value of the reference specimen from the mRNA representative value of the second and subsequent specimens as a correction coefficient, the correction means obtains a small RNA expression level in the second and subsequent specimens. The apparatus performing the correction by subtracting a correction coefficient from the logarithmic value of the measured value.

The apparatus according to claim 7, wherein the expression level of small RNA is comparatively analyzed between two specimens.

The apparatus according to claim 7 or 8 , wherein the representative value is an average value calculated as a logarithmic value of the mRNA expression level measurement value.

To compare and analyze small RNA expression levels among multiple samples,
A storage means for storing a plurality of small RNA expression levels and a plurality of mRNA expression levels measured simultaneously for each of a plurality of samples,
For each specimen, representative value acquisition means for acquiring a representative value represented by a logarithmic value from the measured value of the mRNA expression level,
An input means for inputting a first sample designated as a “reference sample” among a plurality of samples;
Correction coefficient acquisition means for acquiring the difference between the mRNA representative value of the reference specimen and the mRNA representative values of the remaining second and subsequent specimens as a correction coefficient for the second and subsequent specimens,
Correction means for correcting the small RNA expression level measured in the second and subsequent samples using the correction coefficients for the second and subsequent samples acquired by the correction coefficient acquisition means, and the small RNA of the reference sample A program for outputting the comparison result between the expression level and the corrected small RNA expression level in the second and subsequent samples, and functioning as an output means,
When the correction coefficient acquisition unit acquires a value obtained by subtracting the mRNA representative value of the second and subsequent samples from the mRNA representative value of the reference sample as the correction coefficient, the correction unit obtains the small RNA expression level in the second and subsequent samples. The correction is performed by adding a correction coefficient to the logarithmic value of the measured value,
When the correction coefficient acquisition unit acquires a value obtained by subtracting the mRNA representative value of the reference specimen from the mRNA representative value of the second and subsequent specimens as a correction coefficient, the correction means obtains a small RNA expression level in the second and subsequent specimens. The program for performing the correction by subtracting a correction coefficient from a logarithmic value of a measured value.

To compare and analyze small RNA expression levels among multiple samples,
A storage means for storing a plurality of small RNA expression levels and a plurality of mRNA expression levels measured simultaneously for each of a plurality of samples,
For each specimen, representative value acquisition means for acquiring a representative value represented by a logarithmic value from the measured value of the mRNA expression level,
A reference sample selection means for selecting a first sample as a “reference sample” from a plurality of samples;
Correction coefficient acquisition means for acquiring the difference between the mRNA representative value of the reference specimen and the mRNA representative values of the remaining second and subsequent specimens as a correction coefficient for the second and subsequent specimens,
Correction means for correcting the small RNA expression level measured in the second and subsequent samples using the correction coefficients for the second and subsequent samples acquired by the correction coefficient acquisition means, and the small RNA of the reference sample A program for outputting the comparison result between the expression level and the corrected small RNA expression level in the second and subsequent samples, and functioning as an output means,
When the correction coefficient acquisition unit acquires a value obtained by subtracting the mRNA representative value of the second and subsequent samples from the mRNA representative value of the reference sample as the correction coefficient, the correction unit obtains the small RNA expression level in the second and subsequent samples. The correction is performed by adding a correction coefficient to the logarithmic value of the measured value,
When the correction coefficient acquisition unit acquires a value obtained by subtracting the mRNA representative value of the reference specimen from the mRNA representative value of the second and subsequent specimens as a correction coefficient, the correction means obtains a small RNA expression level in the second and subsequent specimens. Performing the correction by subtracting the correction coefficient from the logarithm of the measured value,
The program.

The program according to claim 10 or 11, wherein the expression level of small RNA is comparatively analyzed between two specimens.

The program according to any one of claims 10 to 12 , wherein the representative value is an average value calculated as a logarithmic value of an mRNA expression level measurement value.

The computer-readable recording medium which recorded the program of any one of Claims 10 thru | or 13 .

A chip for small RNA expression analysis, comprising a support on which a plurality of small RNA capture probes and a plurality of mRNA capture probes are immobilized.