酶标板上基于竞争杂交反应的几种miRNA序列的比色检测

Multiplexed miRNA detection is of great importance for early diagnosis of cancers. In this study, a colorimetric method based on the competitive hybridization reaction has been developed in which the biotinylated miRNA (biotin-miRNA) and target miRNA compete for the immobilized oligonucleotide (ODN) probe. Targets miR-182, miR-185 and miR-381 were analyzed using the present method. Relying on the specific interaction between avidin and biotin, complex of avidin-poly-horseradish peroxidase (SA-poly-HRP) was introduced onto the wells of the ELISA plates. The incorporated HRP could catalyze the chromogenic reaction of 3, 3’, 5, 5’-tetramethylbenzidine and H2O2. The method is highly selective, and could determine target miRNAs with concentrations as low as 10 fmol/L, thus holding great potential for clinical miRNA assay.