Colorectal cancer metastasis detection primer and kit based on serum exosomal microRNAs

Abstract

The invention discloses a colorectal cancer metastasis detection primer and a colorectal cancer metastasis detection kit based on serum exosomal microRNAs. According to the primer and the kit, the serum exosomal microRNAs differential expression profiles of colorectal cancer patients suffering from lymphatic metastasis and colorectal cancer patients not suffering from lymphatic metastasis are screened by adopting a second-generation high-throughput sequencing technology, then miR-483-5p with the most obvious up-regulation expression and miR-451a with the obvious down-regulation expression are subjected to large sample verification, and the result shows that miR-483-5p and miR-451a have the good identification and diagnostic efficiencies for colorectal cancer lymphatic metastasis. Meanwhile, combined miR-191-5p and U6 are taken as reference genes for carrying out relative quantification analysis on miR-483-5p and miR-451a in a sample, so that the data accuracy is improved. According to the primer and the kit, the amplification efficiency (E) is calculated according to the formula E=10<(-1 / Slope)>-1, the expression quantities of miR-483-5p and miR-451a relative to miR-191-5p and U6 are calculated according to the formula Q=(E+1)<-delta Cq>, so that the restriction that in the traditional 2<-delta delta Cq> method, the PCR amplification efficiency needs to be 100% is broken through, and the result analysis is relatively reliable.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and relates to a special primer and a kit for detecting colorectal cancer metastasis-related serum exosomal microRNAs, and a method for detecting the expression level of colorectal cancer serum exosomal microRNAs. Background technique [0002] Colorectal cancer is a malignant tumor with high morbidity and mortality. Metastasis is the main cause of treatment failure and death in patients with rectal cancer. Studies have shown that more than half of colorectal cancer patients will eventually metastasize, resulting in tumor recurrence after surgery. From the perspective of clinical treatment effects, accurate prediction of metastasis or high-risk groups of metastasis, and active preventive treatment measures are important links to reduce the recurrence and metastasis of colorectal cancer and improve the prognosis. At present, there are many clinical methods for detecting colorec...

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Problems solved by technology

Coupled with the great heterogeneity among different tumors, exosomal microRNAs related to the metastasis of other tumors have little reference for the detection of colorectal cancer metastasis

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