Detection kit and detection method for pkd1 gene mutation

Abstract

The invention provides a primer set, a kit and a detection reaction system for detecting PKD1 gene mutation through the long fragment PCR and high-throughput sequencing technology, a non-diagnosis-purpose method for external PKD1 gene mutation detection, a non-diagnosis-purpose method for external PKD1 gene analysis and a method for detecting new mutation sites on the PKD1 gene. According to the method, the primer set is used for carrying out long fragment PCR amplification on the PKD1 gene of a sample, and detecting or analyzing is carried out through high-throughput sequencing. The autosomal dominant genetic polycystic nephrosis (adult polycystic nephrosis) can be diagnosed through the assistance of a detection result, previous unknown new mutation on a plurality of PKD1 real genes can be obtained and supplied to doctors or researchers so that the relevance between the mutation and the adult polycystic nephrosis can be studied.

Description

technical field [0001] The invention relates to the technical field of medical molecular biology detection, in particular to a primer set, a kit for detecting PKD1 gene mutations related to hereditary polycystic kidney disease, and a method for detecting PKD1 gene mutations using the primer set and kit. Background technique [0002] 1. Disease and genetic background [0003] Autosomal dominant polycystic kidney disease (adult polycystic kidney disease, APKD, also known as adult polycystic kidney disease) is a common congenital disease. Age-onset is a common hereditary multiple cystic kidney disease with an incidence of about 1 / 1,000-1 / 400, accounting for about 5% of end-stage renal disease. The disease slowly forms cysts and enlargements in the kidneys in middle age, eventually leading to kidney failure. The diagnosis of autosomal dominant polycystic kidney disease mainly depends on imaging or molecular genetic testing. For imaging detection, the sensitivity of detection ...

AI Technical Summary

Problems solved by technology

However, the applicant found that in the method disclosed in this article, the interference of pseudogenes cannot be effectively eliminated after the amplification of long fragments of some primers, and the stability of the primers is still not ideal, and it can only be completed after three times of PCR reaction conditions. Amplification of the entire gene takes longer and is less efficient

This method uses the Illumina Miseq sequencer for detection, which takes 5 to 10 days to complete the sequencing, and the efficiency is low

Moreover, because the GC content of exon 1 sequence of PKD1 gene is greater than 85%, the quality of exon 1 sequencing is poor

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