Multi-nucleic-acid chromatography rapid detecting method and application

Abstract

The invention provides a multi-nucleic-acid chromatography rapid detecting method and an application. The RT-LAMP technology, the nucleic-acid probe hybridization technology and the colloid-gold chromatography technology are combined, and rapid detection of multiple LAMP products is achieved. Colloidal gold test paper is composed of a sample pad, a gold-mark combining pad, a nitrocellulose coating membrane and a water absorbing pad, and the result is judged according to the color developing condition of a testing line and a quality control line. According to the research, the nucleic-acid probe hybridization technology and the colloid-gold chromatography technology are combined, and the multiple LAMP reaction products are detected with the colloid-gold nucleic-acid chromatography method; as expensive special instruments and devices are not required in the detection process, detecting cost is reduced, detecting efficiency is improved, and the multi-nucleic-acid chromatography rapid detecting method is suitable for primary care health units and early screening of HIV and HCV of a large number of persons.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid detection, in particular, the invention relates to a rapid detection method and application of multiple nucleic acid chromatography. Background technique [0002] With the continuous development and improvement of molecular biology and genomics, nucleic acid amplification technology has been gradually applied to the detection of many pathogens. LAMP technology: loop-mediated isothermal amplification (loop-mediated isothermal amplification, LAMP) technology, established by Notomi et al. in 2000, its principle is to use the BstDNA polymerase with strand displacement activity and design for 6 different regions of the target gene 4 specific primers are used to amplify the target gene at a constant temperature of 60-65°C. The entire detection reaction takes only 60 minutes, and the amplification time can be further shortened to 40 minutes in the presence of loop primers. Its reaction sensitivity...

AI Technical Summary

Problems solved by technology

[0003] Due to the complexity of the LAMP reaction system, the amplification products are a mixture of DNA fragments of a series of stem-loop structures and polycyclic cauliflower structures of different sizes, and it is impossible to distinguish the products by directly comparing the molecular weights of the electrophoretic bands.

At present, the detection of LAMP products adopts visual observation and turbidity method to detect the white precipitate of magnesium pyrophosphate, or adds magnesium ion indicator hydroxynaphthol blue to the reaction system or adds fluorescent dye to the final product, and judges by observing the color change before and after the reaction As a result, not only the fluorescent reagents will cause harm to the human body during the detection by the fluorescence method, but also lead to the occurrence of false positives, which limits the popularization and use of the LMAP technology, and the multiple LAMP method is rarely reported.

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