DNA library for detection and diagnosis of polycystic kidney causing genes and application thereof

A DNA library and pathogenic gene technology, applied in the field of DNA library, can solve the problems of inability to accurately distinguish specific types of polycystic kidney disease, inability to meet the timeliness of multi-sample detection, and inability to accurately diagnose young patients, so as to avoid recurrence. Risk of polycystic kidney disease, early occultation, and improved survival

Active Publication Date: 2016-05-04
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, imaging tests are not specific and cannot accurately distinguish the specific types of polycystic kidney disease
Accurate diagnosis also cannot be made in young patients who do not show clinical symptoms at an early stage
Therefore, at present, the combination of clinical symptoms and imaging examinations alone cannot accurately distinguish the various polycystic kidney diseases

Method used

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  • DNA library for detection and diagnosis of polycystic kidney causing genes and application thereof
  • DNA library for detection and diagnosis of polycystic kidney causing genes and application thereof
  • DNA library for detection and diagnosis of polycystic kidney causing genes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] 1. Reagents used in the method:

[0054] Ion AmpliSeq TM LibraryKit2.0, IonPGM TM TemplateOT2200Kitv3, Ion Sequencing200Kitv2, IonXpressBarcodeAdaptors1-16Kit, Ion318 TM ChipKitv2

[0055] 2. Specimen collection and preservation

[0056] (1) Specimen collection: The specimen is the peripheral blood of the patient. Blood is 5ml of venous blood taken routinely, treated with EDTA anticoagulant.

[0057] (2) Storage: It can be detected immediately, stored at 4°C for one week, and stored at -80°C for more than one week.

[0058] 3. Detection steps and result analysis:

[0059] (1) Extraction of genomic DNA of specimens: DNA extraction of specimens was carried out according to the operation instructions of the blood DNA extraction kit of Tiangen Biochemical Technology (Beijing) Co., Ltd.

[0060] (2) Super multiplex PCR amplification and library building of the target detection area: the whole exons of the 6 genes involved in the present invention are used as the det...

Embodiment 2

[0099] In the present invention, the primer pool contains 3 polycystic kidney disease-causing genes newly discovered by the inventor, as shown in the following table:

[0100] serial number

gene name

OMIM number

Associated disease

1

TSC1

605284

autosomal dominant polycystic kidney disease

2

TSC2

191092

autosomal dominant polycystic kidney disease

3

DKK3

605416

autosomal dominant polycystic kidney disease

[0101] The newly discovered 3 polycystic kidney disease genes come from the research accumulation and family investigation of the inventors over the years. The inventor first searches the known polycystic kidney disease-causing genes through the KEGG signaling pathway and the homologous gene library to find the homologous genes of the known polycystic kidney disease genes and the key genes on the same pathogenic pathway. Then, targeted high-throughput sequencing of candidate disease-causing gen...

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PUM

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Abstract

The invention discloses a DNA library for detection of polycystic kidney causing gene mutation by means of the targeted high-flux semiconductor sequencing technology and application thereof. Specifically, a primer pond is designed according to six polycystic kidney causing genes, sample genome DNA is subjected to super-multiple PCR amplification, the amplified product is sequenced by means of the high-flux semiconductor sequencing technology, causing mutation is found, genetic causes of polycystic kidneys are clarified, genetic and molecular biological theoretical bases are provided for clinical diagnosis, support is provided for donor identification in a later-period transplanting operation on a polycystic kidney patient, and a basis is provided for selective good birth and good care by the polycystic kidney patient. The DNA library has the advantages of being accurate, rapid, flexible and low in cost. Through the detection region of the six genes related to the DNA library, autosomal dominant polycystic kidneys and autosomal recessive polycystic kidneys can be detected, and the DNA library has a significant meaning and clinical value for diagnosis and identification of polycystic kidneys.

Description

technical field [0001] The invention relates to a DNA library for detecting and diagnosing polycystic kidney disease-causing genes by targeted high-throughput semiconductor sequencing technology and its application. Specifically, according to the polycystic kidney disease-causing gene, design supermultiplex PCR primers that can cover the exons and adjacent regions of the above genes, and perform supermultiplex PCR amplification on the sample genomic DNA, and the amplified products are analyzed using high-throughput sequencing technology. Sequencing, looking for pathogenic mutations, clarifying the genetic etiology of polycystic kidney disease, providing a theoretical basis for genetics and molecular biology for clinical diagnosis, providing support for the identification of donors for late-stage transplantation in patients with polycystic kidney disease, and conducting clinical trials for patients with polycystic kidney disease Selective prenatal and postnatal care provides th...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/10C40B50/06
CPCC12N15/1093C12Q1/6869C12Q1/6883C12Q2600/156C12Q2600/16C40B50/06C12Q2531/113C12Q2525/191C12Q2537/143C12Q2535/122
Inventor 汪道文吕永曼李宗哲
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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