Nucleic acid detection method and test strip

Abstract

The invention relates to the field of biotechnology and discloses a nucleic acid detection method and a test strip. According to the method, an upstream primer with tag sequence and a downstream primer with a first counterpart are designed according to nucleic acid to be detected; nucleic acid to be detected undergoes PCR amplification; an amplification product undergoes denaturation and the denaturation product is combined with a colloidal gold-coated second counterpart; the combination product is then respectively combined with a detection line capable of capturing target detection objects; and result interpretation is carried out according to developing condition of the detection line. According to the invention, the target nucleic acid is captured through the tag sequence and anti-tag sequence, and the target nucleic acid can be specifically detected at room temperature. By the tag sequence and anti-tag sequence, the problem that the antigen-antibody types in the traditional antigen-antibody colloidal gold technology are few is overcome, and parallel detection of various target detection objects can be realized. The detection method of the invention is simple to operate, can be adopted to read results by naked eyes, is time-saving and labor-saving, and is more beneficial to practical application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a nucleic acid detection method and a test strip. Background technique [0002] In recent years, nucleic acid detection has played an important role in the further study of genetic diseases, the determination of cancerous genes and the screening of drug resistance. For example, single nucleotide polymorphism (Single Nucleotide Polymorphism, SNP), which is a change in the DNA sequence caused by a change in a single nucleotide - A, T, C or G, causes chromosomal changes among species including humans. Genome diversity. The accurate detection of the nucleic acid of the SNP site mutation provides a reference basis for further diagnosis of the disease. Therefore, the detection of nucleic acid mutations is of great significance in the field of life sciences, and the research on nucleic acid detection methods has become one of the research hotspots in the field of life sciences. [0003] ...

AI Technical Summary

Problems solved by technology

However, this technology requires the use of radioactive isotopes as labels, which endangers the safety of researchers. Moreover, the detection cycle of molecular imprinting is long and the operation is complicated;

[0005] 2) Direct sequencing method, which is costly and complicated to operate. For samples with a mutation ratio of less than 15% to 20%, it is difficult to detect by using the sequencing method. Moreover, only one sequence can be detected in one sequencing process. Base composition, for multiple sequences, they need to be sequenced separately, which is time-consuming and labor-intensive;

[0006] 3) Real-time fluorescent quantitative PCR. Fluorescent quantitative PCR technology has the characteristics of high sensitivity, strong specificity, and high degree of automation, but it also has the disadvantages of easy sample contamination and high false positive rate, and can only detect one mutation type at a time;

[0007] 4) Electrophoresis technology, including agarose gel electrophoresis and polyacrylamide gel electrophoresis, etc., is simple to operate and has a short detection period, but electrophoresis technology can only distinguish nucleic acid sequences of different lengths, and which mutation at which site specifically occurs , the determination of the specific composition of the nucleic acid sequence, etc., cannot be known;

[0008] 5) DNA chip technology, this method is fast, simple, and highly automated. However, whether it is a solid-phase chip or a liquid-phase chip, an instrument is required to interpret the results, and the detection instrument is expensive and the operation procedure of the instrument is complicated, so it cannot be used on a large scale. promotion and application;

[0009] 6) Colloidal gold technology, due to its unique optical, electrical properties and biological affinity effects, colloidal gold technology is currently widely used in the fields of catalysis, sensors and DNA analysis. Combining with chromatography technology, you only need to insert the sample liquid when using it, and you can make accurate results in a few minutes. At present, the colloidal gold detection DNA products on the market mainly use DNA antigens to coat the DNA to form the antigen to be tested. , by using the gold-labeled antibody to label the antigen to be tested, the monoclonal antibody is coated on the detection line, and the antibody-antigen to be tested-gold-labeled antibody is formed at the detection line by using the immune reaction principle of antigen-antibody specific binding Complex, anti-gold-labeled antibody-gold-labeled antibody complex is formed at the control line to detect target DNA. In this technology, the combination of various antigens and antibodies is achieved through mutual recognition between antigenic determinants. Different antigens The spatial configuration of antibodies inevitably affects their mutual recognition and binding, thereby affecting the signal strength of the detection, and non-specific binding also makes the false positive rate of this technique high

Images