Application of Asymmetric Multicolor Fluorescent Hairpin Probe Chain Reaction in Pathogen Detection

Abstract

The invention relates to application of an asymmetric multicolor fluorescence hairpin probe chain reaction in pathogenic bacterium DNA (deoxyribonucleic acid) detection, belonging to the field of biotechnological detection. In the invention, in combination with the asymmetric hairpin probe chain reaction technology and the FRET (fluorescence resonance energy transfer) technology, a pathogenic bacterium detection technology based on the asymmetric multicolor fluorescence hairpin probe chain reaction technology is established. If no target sequence exists in the system, the hairpin probe is in a sub-stable state, two groups form an FRET pair, and no fluorescence signal is generated; when the target sequence is added to the reaction system to trigger the chain polymerization reaction, the hairpin structure is opened, and a fluorescence signal is generated; and due to linear amplification, the strength of the fluorescence signal is directly proportional to the copy number of the target sequence. In the method provided by the invention, by introducing the high-sensitivity multicolor FRET technology into the chain reaction, a simple, quick and sensitive pathogenic bacterium DNA direct detection method is established, and the shortcoming of lower sensitivity of the agarose gel electrophoresis observation detection result in the chain reaction of the traditional asymmetric hairpin probe is overcome.

Description

technical field [0001] The invention relates to a biological reaction technology, in particular to the application in the detection of pathogenic bacteria based on asymmetric multicolor fluorescent hairpin probe chain reaction. Background technique [0002] The detection of molecular diagnostic methods based on the characteristics of bacterial nucleic acids has brought new opportunities for the rapid detection of clinical pathogenic microorganisms. Nucleic acid molecular solid-phase hybridization method, DNA sequencing method, capillary electrophoresis analysis, gene chip technology and other detection methods have been successively established, and the bacterial molecular detection method has been raised to a new stage. However, most of these molecular diagnostic methods require special large-scale instruments. The technology is difficult, and it is difficult to popularize and apply it in clinical practice. Therefore, in view of the above problems, if a fast, specific and ...

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Problems solved by technology

Nucleic acid molecular solid-phase hybridization method, DNA sequencing method, capillary electrophoresis analysis, gene chip technology and other detection methods have been successively established, and the bacterial molecular detection method has been raised to a new stage. However, most of these molecular diagnostic methods require special large-scale instruments. The technology is difficult and it is difficult to popularize and apply it in clinical practice

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