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Method for standardizing ncRNA detection result

A technology for detecting and comparing results, which is applied in biochemical equipment and methods, microbe measurement/inspection, etc., to achieve the effects of avoiding operational errors, significant differences, and simple reaction systems

Active Publication Date: 2017-08-29
迪可定(上海)生物科技有限公司
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Problems solved by technology

However, no specific methodological studies mention ratio-based methods applied to cycle ncRNA-sequencing and RT-qPCR data normalization

Method used

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  • Method for standardizing ncRNA detection result
  • Method for standardizing ncRNA detection result
  • Method for standardizing ncRNA detection result

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Data collection and preprocessing

[0055] The ncRNAs expression profile data and reverse transcription real-time PCR quantitative data of 50 cases of diagnosed early lung adenocarcinoma and 29 cases of normal people were obtained from Peking University People's Hospital. Calculate the ratio of sequencing and RT-qPCR data for any two ncRNAs.

[0056] For RT-qPCR data, the calculation of the expression level of the ratio of 2 small molecule ncRNAs (ncRNA1 / ncRNA2) was performed using the comparative CT method (2 -ΔCT ), for the same sample, ΔCT=CT ncRNA1–CT ncRNA2. After the plasma ncRNA concentration was log2-transformed, we used SPSS20.0 software to perform an unpaired T-test to compare the average ncRNA ratio between the adenocarcinoma group, benign patients, and normal controls, with a significant p-value set at 0.05. Chi-square test was performed with SPSS 20.0 software to compare the distribution of gender, race and cancer stage of samples in the training and vali...

Embodiment 2

[0058] Ratio-based approach to normalize circulating ncRNA profile data independent of any internal or external standard controls

[0059] Because neither exogenous nor endogenous controls are reliable for normalization of circulating ncRNA profile data, we tested a ratio-based approach for normalization of circulating ncRNA profile data. First, we calculate the ratio of any two ncRNAs in the same sample, and then compare the ratio of expression levels in different groups instead of comparing the expression level of a single ncRNA. This ensures simultaneous measurements in the same sample under the same conditions, where the same conditions include the same acquisition method, the same storage and the same extraction, PCR or sequencing procedures. This ensures that the relative expression levels of the ratio of the 2 ncRNAs will directly reflect the true value of the comparison. Take miRNA and internal control (IC) as an example (Table 1).

[0060] Table 1 Rates based on nor...

Embodiment 3

[0066] Mathematical logic proof of ratio-based normalization method for miRNA data correction, confirming that ratio-based data normalization method is mathematically logically true

[0067] In order to prove mathematically that the ratio of 2 ncRNAs (such as different miRNAs) in the same sample can reflect the real biological value of 2 ncRNAs. Two assumptions for normalization methods based on endogenous or exogenous controls. That is, first, it is assumed that the miRNA to be tested and the internal control in the same sample are affected by the same system factor; second, it is assumed that the real internal control value is the same in different samples. Ratio-based methods simply assume that different miRNAs in the same sample have the same systematic factor.

[0068] We use miRNA as an example in the proof of mathematical logic, the ultimate goal is to try to find the biologically true miRNA value (trumiRNA), however, usually the observed miRNA (OBSmiRNA) value we get ...

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Abstract

The invention relates to a method for standardizing a detection result of ncRNA in a biological sample. The method is characterized in that the ratio of two ncRNAs is used as a detection classifying index in result judgment; in addition, the method is not independent on any inner source contrast or outer source contrast. Compared with an inner source contrast standardizing method and an outer source contrast standardizing method, the method has the advantage that the logical authenticity of the ncRNA detection result can be improved. The invention specifically provides a method for selecting ncRNA related to lung adenocarcinoma.

Description

technical field [0001] The invention belongs to the field of biological detection methods, in particular to a method for processing and analyzing biological quantitative detection results, more specifically to a method for standardizing the detection results of ncRNA in biological samples based on ratios, and in particular to a lung A method for normalizing the detection results of adenocarcinoma-associated ncRNAs to improve their logical authenticity. Background technique [0002] Lung cancer is a common disease with the most heterogeneity, and it is the number one killer of male cancer. In addition, lung cancer is prone to regional lymph node and distant organ metastasis. Every year, new cases of lung cancer account for 17% of all new cancers in the world, and the number of deaths accounts for 23% of cancer deaths. Lung cancer is the most frequently detected cancer and the leading cause of cancer death in China, with the largest number of new cases and the highest number ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6886C12Q2600/158C12Q2600/178C12Q2525/207C12Q2537/165
Inventor 邓友平王红卫
Owner 迪可定(上海)生物科技有限公司
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