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Nucleic acid detecting method

A detection method and nucleic acid technology, applied in the field of nucleic acid detection, can solve the problems of long detection time, many samples, difficult clinical application, etc., and achieve the effects of simple operation, high sensitivity, and conducive to large-scale popularization and use.

Active Publication Date: 2013-12-04
XIAMEN UNIV
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AI Technical Summary

Problems solved by technology

This type of method takes a long time to detect and requires a large number of samples, making it difficult to achieve large-scale clinical application

Method used

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Embodiment 1

[0058] The preparation of embodiment 1 human papillomavirus (HPV) type 6, type 11, type 16, type 18 rapid typing detection test strip

[0059] In this embodiment, four genotypes of HPV6, HPV11, HPV16, and HPV18 are used as detection objects, and the method provided by the present invention is used to realize rapid typing detection of the four genotypes. The specific implementation steps are as follows:

[0060] 1. Design of PCR amplification primers

[0061] First of all, it is necessary to use software, such as Primer 5.0 and BLAST, to design primers by comparing the sequences of the samples to be tested and according to the principle of LATE-PCR primer design, and then optimize the amplification conditions such as the amount of primers and annealing temperature, and follow-up. Nucleic acid hybridization verification, screening and obtaining single-stranded nucleic acid amplification system with high amplification efficiency and specificity. This system can efficiently ampli...

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Abstract

The invention relates to a technology for detecting nucleic acid and provides a nucleic acid detecting method having the advantages of high sensitivity, high accuracy, low cost, simple operation and quickness, and a nucleic acid film chromatographic hybrid strip thereof. The nucleic acid film chromatographic hybrid strip comprises a sampling pad, target nucleic acid sequence capturing probes, a chromatographic film, a water absorbent pad and a back lining, wherein the chromatographic film is bonded to the middle of the back lining; the sampling pad and the water absorbent pad are bonded to both ends of the chromatographic film respectively; and the target nucleic acid sequence capturing probes are fixed on the chromatographic film between the sampling pad and the water absorbent pad in a straight line. In the method, the colored fluorescence marking probes and the film chromatography technology are combined, the type of a template of a sample can be differentiated by two parameters, namely fluorescence and a physical position, so that the quick genotyping detection of nucleic acid is realized. The nucleic acid detecting method is high in sensitivity, and the detection sensitivity reaches 101 copies / reaction. The nucleic acid detecting method is high in specificity, the high specificity are jointly guaranteed by the primers, detection probes and capturing probes, so that the detection result is more stable and accurate.

Description

technical field [0001] The invention relates to nucleic acid detection technology, in particular to a nucleic acid detection method based on a fluorescent probe. Background technique [0002] At present, there are more and more methods for genotyping detection at home and abroad, such as nucleotide sequence analysis, restriction fragment length polymorphism (RFLP), type-specific primer method and type-specific probe hybridization. Although the sequencing method is the most accurate, it requires high technical requirements and is expensive. RFLP has high sensitivity, but the restriction site is easily affected by gene variation, and in case of mixed infection or incomplete digestion, complex bands will appear, which will affect the judgment of the type result. Type-specific primer polymerase chain reaction (SSP-PCR) requires multiple tube reactions, which is inconvenient to operate and relatively high in cost. The type-specific probe hybridization method is more and more wi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/70C12R1/93
Inventor 李庆阁许晔刘英华夏晓虎
Owner XIAMEN UNIV
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