Constant-temperature amplification method of double-labeled nucleic acid and detection test strip

Abstract

The invention discloses a constant-temperature amplification method of double-labeled nucleic acid and a test strip. The constant-temperature amplification method comprises the following steps of: adding labeled primers into an LAMP (loop-mediated isothermal amplification) reaction system for constant-temperature amplification by firstly synthesizing inner primers, outer primers and ring primers and using biotin and fluorescein to label the inner primers or / and the ring primers, and obtaining a constant-temperature amplification product of the double-labeled nucleic acid. The test strip comprises a supporting layer and a nucleic acid amplification product adsorbing layer which is pasted on the supporting layer; and the biotin affinity protein and fluorescein isothiocyanate antibody are used for printing a detection mark and a control mark on a cellulose layer. In the test strip disclosed by the invention, the detection specificity is strong, the sensitivity is high, the speed is high,not only can the LAMP product be intuitively detected, but also the defect that the specificity of the detection test paper of the colloidal gold is lower can be overcome. The invention can be widelyused for detection of specific genes in various pathogenic microorganisms or pathogens.

Description

technical field [0001] The invention relates to a ring-mediated isothermal amplification method and a test strip, in particular to a double-labeled nucleic acid amplification method and a rapid detection test strip. Background technique [0002] With the development of modern molecular biology and molecular technology, many nucleic acid amplification techniques have been researched and developed. Among them, the nucleic acid loop-mediated isothermal amplification technology (loop-mediated isothermal amplification, LAMP), due to its strong specificity, high sensitivity, and fast reaction speed, has a tendency to replace PCR in the field of nucleic acid-specific amplification and detection. It has become one of the research hotspots in the field of life sciences. The main principle of nucleic acid circle-mediated amplification is based on designing 2 pairs of specific primers for the 6 regions of the target sequence, using a strand-displacing DNA polymerase (Bst DNA polymeras...

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Problems solved by technology

[0003] At present, electrophoresis and turbidimetry are often used to detect LAMP products, which requires the use of highly carcinogenic fluorescent dye ethidium bromide, which not only endangers the health of operators, but also cannot rule out the problem of false positives, which limits the promotion and use of LAMP technology.

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