Probe, primer, detection reagent kit and detection method for detecting hereditary hearing impairment on basis of floating-bead array system

A technology of hereditary deafness and detection kit, which is applied in the field of molecular biology and can solve problems such as addressing

Active Publication Date: 2017-03-08
广东辉锦创兴生物医学科技有限公司 +1
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Problems solved by technology

The probes on the solid plane are addressed through the orderly positional relationship between rows and columns; while in the microbead suspension array technology, the microbeads are flowing, so naturally they cannot be addressed by position. so only by encoding the bead itself

Method used

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  • Probe, primer, detection reagent kit and detection method for detecting hereditary hearing impairment on basis of floating-bead array system
  • Probe, primer, detection reagent kit and detection method for detecting hereditary hearing impairment on basis of floating-bead array system
  • Probe, primer, detection reagent kit and detection method for detecting hereditary hearing impairment on basis of floating-bead array system

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Embodiment

[0083] The present invention is based on the primers and probes of the suspended microbead array system for detecting hereditary deafness, and can be compatible with two kinds of suspended microbead array systems to detect genetic mutations of hereditary deafness. , 299_300delAT, 35delG, 176_191del16, 512insAACG, 109G>A); 1 locus of GJB3 gene (538C>T); 11 loci of SLC26A4 gene (IVS7-2A>G, 2168A>G, 1174A>T, 1229C>T , 1226G>A, 1975G>C, 2027T>A, IVS15+5G>A, 1693insA, 754T>C, 2086C>T); the mitochondrial 12S rRNA gene 2 sites rRNA (1494C>T, 1555A>G) totaled 20 Detection of mutation sites.

[0084] 1. Multiplex PCR primer design

[0085] In order to realize the detection of 20 genetic mutation sites of hereditary deafness, a total of 20 PCR amplifications are required. The PCR primer sequences corresponding to these 20 mutation sites are shown in Table 1, wherein a universal primer (deaf-Tag) is added to the 5' end of the reverse primer (R). Biotin modification was performed at th...

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Abstract

The invention discloses a probe, a primer, a detection reagent kit and a detection method for detecting hereditary hearing impairment on the basis of a floating-bead array system. The probe used for mutation of detecting hereditary hearing impairment genes is coupled and mixed with microbeads of different codes to acquire a floating-bead array; the specific primer is used for performing multiple PCR (polymerase chain reaction) amplification, products of the floating-bead array is subjected to hybridization with the floating-bead array, streptavidin R-phycoerythrin is used for color developing, and detecting results are read through a floating-bead array reading instrument. By the arrangement of the probe and the primer for detecting hereditary hearing impairment on the basis of the floating-bead array system, 20 mutation sites of the four major hereditary hearing impairment genes can be detected simultaneously, detection time can be greatly shortened, and detecting efficiency and accuracy can be improved.

Description

[0001] Technical field: [0002] The invention belongs to the field of molecular biology, and in particular relates to a group of probes, primers, detection kits and detection methods for detecting hereditary deafness based on a suspended microbead array system. [0003] Background technique: [0004] Hearing disability ranks first in my country's disability. According to the "National Total Number of Disabled Persons and the Number of Various Types and Different Levels of Disabled People at the End of 2010" released by the China Disabled Persons' Federation, among the 85.02 million disabled people in my country, 20.54 million people are hearing disabled, accounting for nearly 1 / 4. And the number of newly born deaf children is increasing at a rate of 30,000 cases per year. Genetic factors account for about 60% of the causes of Chinese deafness. For this reason, the detection of hereditary deafness can block the occurrence of most hereditary deafness from the source. [0005]...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 陈敏汪定亮
Owner 广东辉锦创兴生物医学科技有限公司
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