Method and primers for detecting mi ribonucleic acid (miRNA) and application of method

Abstract

The invention discloses a method and primers for detecting mi ribonucleic acid (miRNA) and application of the method. The method comprises the following steps of: performing reverse transcription on the miRNA in a sample by using a reverse transcription primer formed by specific basic groups and Oligo (dT); and performing real-time polymerase chain reaction (PCR) quantitative detection on the miRNA by using a specific forward primer, a general reverse primer and a general probe. The invention has the characteristics that: the sensitivity and the specificity of the method are obviously higher than those of the conventional method; high-throughput analysis can be performed; and the method is simply and quickly operated, is low in cost, and can be widely used for early diagnosis and prediction of critical diseases such as tumors and the like.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method and primers for detecting miRNA and its application in early diagnosis and prognosis of major diseases such as tumors. Background technique [0002] microRNA (miRNA) is a kind of non-coding single-stranded small molecule RNA with a length of about 22 nucleotides, which widely exists in eukaryotic organisms such as animals, plants, and nematodes. miRNA degrades target mRNA or represses its post-transcriptional translation by specifically binding to the 3' untranslated region (3'UTR) of target mRNA, which ultimately leads to a decrease in protein synthesis of the target gene. miRNAs are involved in the regulation of all levels of life activities, including embryonic development, organ formation, cell proliferation, cell apoptosis, cell stress response, stem cell differentiation, endocrine regulation, immune regulation and the occurrence and development of diseases. A large numb...

AI Technical Summary

Problems solved by technology

The stem-loop primer method uses primers with 6 bases at the 3' end that are complementary to the miRNA for reverse transcription of the miRNA. At the same time, the 5' end of the reverse transcription primer contains a stem-loop structure, which can enhance the heterogeneous duplex of miRNA and DNA. strand affinity, and prevent primers from annealing to pri- or pre-miRNA, so in general the specificity of the stem-loop primer method is better than the poly(A) polymerase tailing method; but because the method uses a sequence-specific probe needles, too expensive for high-throughput miRNA analysis

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