Probe set and kit used for detecting related genes of congenital cataract

A congenital cataract and probe set technology, applied in the direction of recombinant DNA technology, DNA / RNA fragments, etc., can solve the problem of not having a relatively comprehensive congenital cataract-related gene region capture probe set, etc., to improve coverage and efficiency, Good specificity and high sensitivity

Inactive Publication Date: 2017-01-04
ZHONGSHAN OPHTHALMIC CENT SUN YAT SEN UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0004] In recent years, with the development of next-generation sequencing technology, the number of reported congenital cataract-related genes has increased day by day, including at least 250 mutation sites in 120 genes. However, there is still no relatively comprehensive congenital cataract-related gene region on the market Capture Probe Combinations and Kits

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  • Probe set and kit used for detecting related genes of congenital cataract
  • Probe set and kit used for detecting related genes of congenital cataract
  • Probe set and kit used for detecting related genes of congenital cataract

Examples

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Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: A probe set for detecting congenital cataract-related genes

[0016] 1. Screening and determination of genes related to congenital cataract

[0017] Using "congenital cataract", "inherited cataract", "pediatric cataract", "infantile cataract", "childhood cataract" as the search terms, in PubMed, Online MendelianInheritance in Man (OMIM) and National Center for Biotechnology Information (NCBI) Search the sub-site for the reported congenital cataract-related genes with different levels of evidence found through family linkage analysis, population association analysis, sporadic case gene detection and animal experiments. Through further screening of clinical phenotype and genetic background, 225 congenital cataract-related genes were finally included for product design and testing.

[0018] The screened congenital cataract-related genes are as follows:

[0019]PEX10(5192),PEX14(5195),EPHA2(1969),HSPG2(3339),POMGNT1(55624),FOXE3(2301),RPE65(6121),CRYZ(1429),DNA...

Embodiment 2

[0081] Example 2: A kit for detecting congenital cataract-related genes

[0082] The kit for detecting congenital cataract-related genes in this embodiment is a kit for molecular genetic detection by detecting mutations in exons of congenital cataract-related genes.

[0083] The kit used to detect congenital cataract-related genes in this embodiment includes: the probe set obtained in Example 1, also includes PCR ion amplification mix (PCR buffer, dNTPs, Taq enzyme, etc.), 96-well plate and 96 Well plate sealing mold and QIAGEN Purification kit (Cat.No.159992), Qiagen PCR kit (Cat.No.28104), QiagenMinElute kit (Cat.No.28004) and Ampure beads XP (Cat.No.4471250), etc.

Embodiment 3

[0084] Example 3: Application of the kit for detecting congenital cataract-related genes

[0085] The genomic DNA that passed the quality inspection was randomly broken into fragments with a length of 180-280bp using a Covaris crusher, and after end repair and A-tailing, adapters were connected to both ends of the fragments to prepare a DNA library. After polling the DNA library with a specific index, perform liquid-phase hybridization with biotin-labeled probes, and then use streptavidin-containing magnetic beads to bind the exon region of the target gene (congenital cataract-related gene) After being captured, the library quality inspection is performed after linear amplification by PCR, and sequencing is performed after passing the standard.

[0086] 1. Sample requirements

[0087] Genomic DNA sample (total DNA sample, OD260 / 280 value between 1.8-2.0, concentration ≧100ng / μL, total amount ≧50μL / sample, and ensure that the DNA is free from contamination and degradation)

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Abstract

The invention discloses a probe set and a kit used for detecting the related genes of a congenital cataract. The probe set comprises a plurality of probe sequences which respectively aim at the exon non-repeated areas of 225 related genes of the congenital cataract. The invention also provides the kit which contains the probe set and is used for regional capture and next generation sequencing. According to the probe set and the kit, the found 225 related genes of the congenital cataract can be comprehensively detected in one time, the coverage rate and the efficiency of molecular diagnosis can be improved, specificity is good, and sensitivity is high, and the probe set and the kit have an important meaning for the prenatal screening, the early diagnosis and the accurate prevention and control of the congenital cataract.

Description

technical field [0001] The invention belongs to the field of gene detection kits, and in particular relates to a probe set and a kit for detecting genes related to congenital cataract. Background technique [0002] Congenital cataract is the opacity of the lens caused by congenital heredity or developmental disorder that exists at birth or gradually forms after birth. Severe visual impairment (best corrected visual acuity is less than 0.3, not less than 0.05) accounts for 50% of the total number of children, and its effective prevention and treatment has always been an international problem that has attracted much attention and needs to be solved urgently. [0003] About 30% of children with congenital cataract have obvious familial inheritance tendency, and there are three inheritance modes, including autosomal dominant inheritance, autosomal recessive inheritance and X-linked inheritance. Of these, autosomal dominant inheritance is the most common form of inheritance. Ba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883
Inventor 刘奕志林浩添吴晓航陈伟蓉
Owner ZHONGSHAN OPHTHALMIC CENT SUN YAT SEN UNIV
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