Method for evaluating quality of body fluid specimen

A body fluid sample and quality technology, which is applied in the field of quality evaluation of body fluid samples, can solve problems such as the degree of decomposition, the time and effort required, and the need for whole genetic data.

Pending Publication Date: 2021-03-16
TORAY IND INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, when measuring the presence of miRNA, the RNA of the short-chain fraction is mostly used, and the long-chain RNA is not contained at this time, so the existing method as described above is not an effective method for measuring the decomposition degree of RNA
It is also possible to measure the degree of decomposition of the RNA used based on the correlation coefficient of the whole gene of the gene expression analysis result, but the data of the whole gene is required, which takes time and effort

Method used

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  • Method for evaluating quality of body fluid specimen
  • Method for evaluating quality of body fluid specimen
  • Method for evaluating quality of body fluid specimen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0179] Selection of reference miRNA capable of detecting degradation in whole blood coagulation

[0180] (Sample preparation for detection of deterioration due to temperature influence)

[0181] The blood of 7 blood collection tubes was collected from 3 healthy people respectively, and in the state of whole blood, one of the 7 tubes was left to stand at room temperature (23°C) for 0.5 hours (this was used as a reference condition), and the remaining 6 tubes were The branch was left still for 6 hours at each temperature of 4°C, 18°C, 20°C, room temperature (23°C), 28°C, and 30°C. After the time was up, centrifugation was performed respectively, and the obtained serum was divided into 300 μL each within 10 minutes after centrifugation, and stored in a freezer at -80°C.

[0182] (sample preparation for detection of deterioration caused by prolonged standing at room temperature)

[0183] The blood of 4 blood collection tubes was collected from 3 healthy people respectively. In ...

Embodiment 2

[0199] Detection of deterioration during coagulation of whole blood with various miRNAs

[0200] The deterioration of the quality of the bodily fluid sample may be determined by combining any two reference miRNAs instead of a single miRNA.

[0201] Existing amounts of hsa-miR-204-3p (SEQ ID NO: 1) and hsa-miR-4730 (SEQ ID NO: 2) in whole blood under the standard conditions of Example 1 and under the condition of standing at 30° C. for 6 hours. The respective presence of these miRNAs under each condition is as follows Figure 5 As shown, the difference in the amount of these two miRNAs under each condition is calculated, then as Figure 6 like that. as table 3 and Figure 5 As shown, hsa-miR-204-3p is an miRNA that shifts to a low value through the degradation of the samples generated in the whole blood state, and hsa-miR-4730 shifts to a high value through the degradation of the samples generated in the whole blood state Of the miRNAs present in undegraded samples, hsa-mi...

Embodiment 3

[0204] Selection of reference miRNA capable of detecting deterioration in serum state

[0205] (Sample preparation for detection of deterioration caused by prolonged standing at 4°C in serum state (modulation 1))

[0206] The blood of 4 blood collection tubes was collected from 3 healthy people respectively, and all were left still at room temperature (23° C.) for 0.5 hour, and then centrifuged to obtain serum. The serum obtained from one mouse was aliquoted into 300 μL each within 10 minutes after centrifugation, and stored in a freezer at -80°C (this was used as a reference condition). The serum obtained from the remaining 3 tubes was left standing at 4°C for 12 hours, 21 hours, and 24 hours. After the time was up, the serum was divided into 300 μL each and stored in a freezer at -80°C.

[0207] (sample preparation for detection of deterioration by standing in serum state (modulation 2))

[0208] Blood in 7 blood collection tubes was collected from 3 healthy people, and a...

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Abstract

Provided is a method in which a reference miRNA is identified that exhibits a change in the amount thereof present in a body fluid specimen depending on changes in the quality of the body fluid specimen, and the quality of the body fluid specimen is evaluated using the amount of the reference miRNA present in the specimen as an index.

Description

technical field [0001] The present invention relates to a method for evaluating the quality of a body fluid sample based on the amount of specific miRNA contained in the body fluid sample. Background technique [0002] miRNA (microRNA) is transcribed from genomic DNA as RNA (precursor) with a hairpin-like structure. This precursor is cleaved by a dsRNA cleavage enzyme (Drosha, Dicer) having a specific enzyme RNase III cleavage activity, becomes double-stranded, and then becomes single-stranded. Also, it is generally believed that one antisense strand is taken up by a protein complex called RISC, which is involved in the translational repression of mRNA. As described above, since miRNAs have different forms at various stages after transcription, it is generally necessary to consider various forms such as hairpin structures, double-stranded structures, and single-stranded structures when miRNAs are detected. miRNA is composed of RNA of 15 to 25 bases, and its existence has b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12M1/00C12Q1/6837G01N33/53C12N15/113
CPCC12Q1/6876C12Q2600/178C12Q2600/166C12Q1/6837C12Q2600/158
Inventor 星野笑美芹泽崇名取一惠
Owner TORAY IND INC
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