Nucleic acid detection method combining RNA amplification with hybrid capture method

Abstract

The invention relates to a nucleic acid detection method combining RNA amplification with a hybrid capture method. The nucleic acid detection method includes the following steps of extracting nucleic acid from a sample or splitting the sample to release the nucleic acid to be detected, conducting transcription and amplification on the nucleic acid to be detected to obtain an RNA product, adding the obtained RNA product together with a DNA probe marked with biotins to a solid phase so that hybridization can be conducted, forming a DNA / RNA heterozygote through hybridization, coating the solid phase with specific antibodies for recognizing the heterozygote or avidin or streptavidin, capturing the heterozygote through the solid phase, and adding avidin or streptavidin or antibodies for recognizing the DNA / RNA heterozygote to the solid phase to achieve obtaining of a nucleic acid signal, wherein the avidin or the streptavidin or the antibodies for recognizing the DNA / RNA heterozygote are marked with enzymes with the secondary amplification capacity. According to the nucleic acid detection method, the inherent pollution problem of the PCR is avoided, RNA molecules can be directly detected, inverse transcription and pre-dissociation of RNA do not need to be conducted, and the operation steps are greatly simplified.

Description

technical field [0001] The invention relates to a nucleic acid detection method combining RNA amplification and hybrid capture method, which belongs to the field of biological detection. Background technique [0002] Polymerase chain reaction technology (PCR) has been widely used in the fields of medicine and molecular biology due to its sensitivity, specificity and rapidity since it came out in 1985, which shows the importance of nucleic acid amplification technology. In recent years, with the rapid development of molecular biology and the extensive application of biophysical techniques, nucleic acid amplification models are also emerging. Direct amplification of target nucleic acid mainly includes polymerase chain reaction (PCR), strand substitution amplification (SDA), ligase chain reaction (LCR) and nucleic acid sequence-dependent amplification (NASBA), etc. High sensitivity. [0003] According to different nucleic acid amplification conditions, nucleic acid amplificat...

AI Technical Summary

Problems solved by technology

This detection method similar to enzyme-linked immunoassay is easy to operate and requires less equipment investment, but there are still deficiencies in sensitivity, specificity and high-throughput detection.

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