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

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...

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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

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