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Nucleic acid quantitative analysis method based on intelligent equipment assistance and application thereof

A quantitative analysis, intelligent equipment technology, applied in biochemical equipment and methods, microbial determination/inspection, instruments, etc., can solve the problems of inability to achieve quantitative analysis, contamination of amplification products, etc., and achieve simple operation and low analysis cost. Effect

Active Publication Date: 2019-09-20
UNIV OF SCI & TECH BEIJING
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Problems solved by technology

[0003] At present, researchers use polymerase chain reaction (PCR), loop-mediated isothermal nucleic acid amplification (LAMP) and strand displacement amplification (SDA) amplification techniques to amplify target DNA / RNA, and use pH indicators to establish A visual nucleic acid amplification product detection system, (BioTechniques, 2015, 58, 59-68.) identifies the "positive or negative" of the target DNA / RNA through the color change before and after the amplification reaction, although a simple and fast target DNA / RNA RNA detection, but not quantitative analysis
Li et al. used PCR, rolling circle amplification (RCA) and SDA amplification combined with a pH indicator to measure the UV-visible spectrum of the amplification product, thereby realizing the quantitative detection of miRNA, (Anal.Chem., 2017, 89, 6631-6636.) However, this still requires additional operation and determination steps, which is likely to cause contamination of the amplification product, and still requires expensive equipment

Method used

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  • Nucleic acid quantitative analysis method based on intelligent equipment assistance and application thereof
  • Nucleic acid quantitative analysis method based on intelligent equipment assistance and application thereof
  • Nucleic acid quantitative analysis method based on intelligent equipment assistance and application thereof

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Embodiment

[0058] Embodiment: BCR-ABL fusion gene mRNA analysis

[0059] Using the molecular markers detected in the clinical diagnosis, treatment and prognosis of chronic myelogenous leukemia (CML), BCR-ABL fusion gene mRNA as the RNA detection model, according to the previously authorized patent (patent number: ZL201610316771.0) to design and synthesize a stem-loop Structural DNA probes SLP-e13 and SLP-a2, the base sequence of the probe SLP-e13 is 5'-CTTCCTTATTGATGGTCAGCTTTATCGTCGTGACTGTTTGTAATAGGACAGAGCCCCGCACTTTCAGTCACGACGAT-3' (provided by Treasure Bioengineering (Dalian) Co., Ltd.);

[0060] The base sequence of the probe SLP-a2 is 5′-CGACAGCAGAGGATTTGTTGTGTGGAAGTGTGAGCGGATTTTCCTCTGCTGTCGTTTTACTGGCCGCTGAAGGGCTT-3′ (provided by Treasure Bioengineering (Dalian) Co., Ltd.), wherein the 5′ end of the probe SLP-a2 is modified with a phosphate group.

[0061] Such as figure 2 As shown, according to the BCR-ABL fusion gene mRNA sequence, two specific stem-loop structure DNA probes SLP-e...

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Abstract

The invention provides a nucleic acid quantitative analysis method based on intelligent equipment assistance and application thereof. According to the method, through designing and constructing a nucleic acid amplification reaction system which is initiated by a nucleic acid object and is sensitive to pH, utilizing a pH indicator cresol red, and ensuring a color changing range between yellow 7.2-pink 8.8, visual quick analysis of the nucleic acid by naked eyes is realized. Based on the visual quick analysis, an intelligent device aided three-primary-color image scanning mode is utilized for realizing quick detection and quantitative analysis of the nucleic acid by intelligent device detecting terminal, wherein the used intelligent device has a photographing function, such as smart phone and panel computer.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, and specifically relates to a nucleic acid quantitative analysis method based on intelligent equipment and its application. The method is to perform nucleic acid amplification reaction in a low buffer system and utilize H + The ions cause the pH change of the system, and the amplification system is monitored by using the color change of the pH indicator. The present invention uses the App in the smart device (mobile phone, tablet, etc.) to read the color information of the nucleic acid amplification reaction system, that is, the three primary colors of the image (red, green, etc.) , blue), establish the relationship between the three primary color values ​​and the nucleic acid concentration, and realize the simple, rapid, high-sensitivity, and high-specificity quantitative analysis of nucleic acids. Background technique [0002] Simple, rapid, highly sensitive, and highly specific nuc...

Claims

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Application Information

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IPC IPC(8): G16B45/00G16B40/00C12Q1/6844
CPCC12Q1/6844G16B40/00G16B45/00C12Q2521/501C12Q2531/119
Inventor 王辉李正平王洪红孙茹艳贾玉婷
Owner UNIV OF SCI & TECH BEIJING
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