Bladder cancer mutation gene specific primer, kit and library construction method

Abstract

The invention discloses a bladder cancer mutation gene specific primer, kit and library construction method. A mode of unilateral enrichment-single molecule specific primer (single primer) is used foramplifying sub enrichment technologies for the first time; the application scope of the primer is expanded, the use amount of an effective template is increased, especially the detection limit for detecting low-frequency mutation samples is greatly improved; and secondly, a single molecule specific primer is composed of specific sequences and sequencing sequences, the specificity and the primer use ratio of multiple PCR can be effectively provided. The bladder cancer mutation gene specific primer, kit and library construction method applies the technology to the field of bladder cancer tumormutation gene detection, prepares corresponding reagents, can detect the mutation condition of tumor genes related bladder cancer efficiently and sensitively, and provides guidance for clinical application.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and in particular relates to a bladder cancer tumor mutation gene-specific primer, kit and library construction method. Background technique [0002] Urinary system tumors mainly include bladder cancer, kidney cancer, urethral cancer, etc., and bladder cancer is the most common urinary system tumor. At present, the examination of bladder cancer mainly relies on imaging examination and pathological examination, etc., but generally expensive and invasive means; urine cytology examination is convenient and non-invasive, but the sensitivity is not very high; cystoscopy is the gold standard for diagnosis of bladder cancer , is also one of the main means of postoperative recurrence monitoring, but it is invasive and may cause related complications; these defects limit the early diagnosis and treatment of bladder cancer, as well as prognosis monitoring. [0003] Researchers continue...

AI Technical Summary

Problems solved by technology

[0004] In traditional gene mutation detection methods, double-sided primers are used for amplification. Because the fragmentation of the region to be detected is random, templates with poor fragment integrity cannot be covered by double-sided primers at the same time, resulting in the loss of trace templates and affecting the detection limit.

There are also unilateral enrichment-strand circling (back-to-back primer) modes and unilateral enrichment-nested PCR primers (nested PCR primers) modes; while the single-measure enrichment-strand circling mode has no single-molecule labeling technology reduction. Noise, single-test enrichment-nested PCR primer mode adopts double primers, which leads to a certain distance between the 3' end of the primer and the site to be detected, and a small amount of template loss, which seriously affects the detection limit

Images