Specific detection primers and detection liquid phase chip for HNF1B gene mutation

Abstract

The present invention discloses a detection liquid phase chip and specific primers for HNF1B gene mutation. The liquid phase chip mainly comprises: ASPE primers comprising a 5' terminal tag sequence and a 3' terminal target gene mutational site-targeted specific primer sequence, wherein the specific primer sequence comprises A12057G site-targeted SEQ ID NO.11, A12057G site-targeted SEQ ID NO.12, C8941T site-targeted SEQ ID NO.13, C8941T site-targeted SEQ ID NO.14, G35118A site-targeted SEQ ID NO.15, G35118A site-targeted SEQ ID NO.16, C3784A site-targeted SEQ ID NO.17, C3784A site-targeted SEQ IDNO.18, and / or G13582A site-targeted SEQ ID NO.19 and G13582A site-targeted SEQ ID NO.20; anti-tag sequence coated microspheres; and amplification primers. According to the present invention, coincidence frequency of the detection results of the detection liquid phase chip and the sequencing method is up to 100%, and single and parallel detection on the wild-type with multiple mutational sites and the mutant-type with multiple mutational sites can be achieved.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for HNF1B gene mutation detection and a liquid phase chip. Background technique [0002] Hepatocyte nuclear factor 1β (hepatocyte nuclear factor1β, HNF1B), also known as transcription factor 2 (transcription factor2, TCF2). The gene is located on chromosome 17 17q12, between 36046433 and 36105095 base pairs. HNF1B gene plays a role in kidney development, regulates the development of embryonic pancreas, and is a transcription factor that plays an important role in the formation of pancreatic cells and the maintenance of blood glucose homeostasis. At present, a large number of studies have shown that HNF1B gene mutation will lead to renal capsule and diabetes syndrome, that is, maturity-onset diabetes of themyoung5 (MODY5), which is manifested as early-onset diabetes, kidney, pancreas and reproductive disease. Abnormal deve...

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Problems solved by technology

[0003] Currently, HNF1B gene mutation detection methods mainly include: Illumina fiber optic microbead chip technology, fluorescence quantitative PCR technology, SNPlex Genotyping System technology and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), although Illumina fiber optic microbead chip technology The technology is a high-throughput detection system with high sensitivity and accuracy, but the degree of automation is low, and there are many manual operations, which are difficult to meet the needs of practical applications. Fluorescence quantitative PCR technology has the characteristics of high sensitivity, strong specificity, and high degree of automation, but There are also the disadvantages of easy sample contamination and high false positive rate, and only one mutation type can be detected at a time. The SNPlexTM System technology has high requirements for sequence specificity of single nucleotide polymorphism sites (SNPs), and cannot be arbitrarily It is difficult to apply to clinical detection and diagnosis, and the operation of this method is relatively complicated, which cannot meet the needs of practical application.

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry is a soft ionization technology that has powerful and mature functions in the detection of protein and other biological macromolecules. However, in the field of nucleic acid detection, due to the particularity of nucleic acid molecules, detection is subject to certain limit

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