Instant detection system and method for nucleic acid of pathogens

Abstract

The invention discloses a real-time detection system and method for nucleic acid of pathogens. The system comprises: a lysis buffer, which is used for carrying out inactivation lysis on a to-be-detected sample to release RNA; a RT premixed solution, which is used for reversely transcribing RNA in a sample into DNA; a RPA premixed solution, which is used for carrying out RPA reaction on the sample treated by the RT premixed solution so as to realize amplification of DNA in the sample; a CRISPR reaction premixed solution, which is used for carrying out specific cleavage reaction on the sample treated by the RPA premixed solution; and colloidal gold test paper, which is used for developing the sample treated by the CRISPR reaction premixed solution so as to confirm whether the sample contains nucleic acid of pathogens or not. According to the above design scheme, a RPA method, a CRISPR method and a colloidal gold method are coupled, so the real-time detection system has the advantages of simplicity, sensitivity and specificity at the same time, does not need instruments and equipment, and is a real-time detection system capable of quickly obtaining a result through simple operation at room temperature.

Description

technical field [0001] The application belongs to the technical field of nucleic acid detection, and in particular relates to a system and method for instant detection of pathogen nucleic acid. Background technique [0002] The current detection method for pathogen nucleic acid is usually reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR), which has high sensitivity and low cross-contamination, and is the most commonly used detection method in front-line hospitals. RT-qPCR technology first reverse-transcribes viral RNA into cDNA, and then performs polymerase chain reaction. Its signal generation generally has two different methods: (1) specific fluorescent labeling method, which requires designing a segment complementary to the target DNA. TaqMan probes have fluorescent groups and quenching groups at both ends of the probe; (2) non-specific fluorescent labeling method, which requires the use of fluorescent dyes such as SYBR Green, which can comb...

AI Technical Summary

Problems solved by technology

In the first method, the fluorescence generation depends on the combination of the probe and the target DNA. The probe needs to be carefully designed. If the template is incomplete or the amplification efficiency is low, false negatives are prone to occur. The second method requires a large amount of amplification of the target DNA. If the sample has DNA contamination and non-specific probe binding, it is very easy to produce false positives

Moreover, RT-qPCR and other common nucleic acid detection techniques rely heavily on a variety of sophisticated equipment, which is not suitable for general public self-test or use in poor or remote areas with limited resources

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