A genetic detection kit for hereditary deafness

A technique for hereditary deafness and genetic testing, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve the problems of few detection sites, complicated operation, time-consuming and laborious, and high cost

CN106811533BActive Publication Date: 2020-10-16亚能生物技术(深圳)有限公司
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2020-10-16

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Abstract

The invention relates to the field of gene detection, in particular to a nucleic acid membrane strip and a kit for hereditary hearing loss gene detection. The kit comprises a PCR reaction liquid, wherein the PCR reaction liquid comprises a PCR reaction liquid A, a PCR reaction liquid B, a PCR reaction liquid C, a PCR reaction liquid D and a PCR reaction liquid E and also comprises corresponding primers and probes. According to the kit, the specific amplification LATE-PCR special primers are designed by use of the sequence of various mutation sites, reported by in literature, and the locked nucleic acid modified ZNATM probes are adopted, so that the least tubes of the reaction liquid can detect the most sites. The PCR reaction liquids A, B, C, D and E in the kit can be tested on the same fluorescent PCR instrument by use of the same amplification program, and the clinical requirement for quick and convenient hearing loss detection is satisfied.
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Description

technical field

[0001] The invention relates to the field of gene detection, in particular to a genetic detection kit for hereditary deafness. Background technique

[0002] At present, the existing deafness gene detection products on the market mainly include Boao's nine hereditary deafness-related gene detection kits (microarray chip method) and 15 hereditary deafness-related gene detection kits (microarray chip method); Kaipu's deafness susceptibility gene detection kit (PCR + diversion hybridization method); Zhongsheng Beikong's four deafness gene detection kit (ARMS-PCR method); Jinan Yingsheng's congenital deafness gene detection kit (fluorescence PCR method), drug-induced deafness gene detection kit (fluorescence PCR method), deafness gene GJB2 235delC detection kit (fluorescence PCR method) and PDS gene mutation detection kit (fluorescence PCR method); Zhihai Bioengineering Co., Ltd. Deafness gene mutation detection kit (fluorescent PCR method). Although there are m...

Examples

Embodiment 1

[0097] The kit of the present invention can simultaneously detect 30 mutation sites in four hotspot genes related to hereditary deafness, that is, GJB2 (35delG, 109G>A, 155delTCTG, 167delT, 176-191del16, 235delC, 299-300delAT, 512insAACG) , GJB3(538C>T, 547G>A), SLC26A4(281C>T, 589G>A, 749T>C, 754T>C, IVS7-2A>G, 1174A>T, 1226G>A, 1229C>T, 1975G> C, 2027T>A, IVS15+5G>A, 2162C>T, 2168A>G), mtDNA (961T>G, 1095T>C, 1494C>T, 1555A>G, 7444G>A, 7445A>G, 12201T>C ).

[0098] 1. Design and screening of primers

[0099] First, use the sequences of various mutation sites reported in the literature to design specific amplification LATE-PCR primers for the purpose of using multiple asymmetric PCR to detect multiple genotypes in the same tube (a total of five tubes), by randomly combining different Primers, optimize the annealing temperature gradient, and finally select a batch of primers with good specificity, which can meet the clinical needs of rapid and convenient detection of deafnes...

Embodiment 2

[0148] The hereditary deafness gene detection kit of the invention adopts LATE-PCR and multicolor fluorescent probe PCR melting curve technology. Design corresponding probes in the mutation region to be detected, and design an upstream primer and a downstream primer on the periphery of the designed probe, use the upstream primer and downstream primer to PCR amplify the fragment containing the region to be detected, and the PCR amplification ends followed by melting curve analysis. Subtract the corresponding Tm value of the standard control in each detection channel from the Tm value of the sample to be tested in each detection channel to obtain the ΔTm value of each detection channel, and judge whether there is a mutation and the type of mutation in the sample to be tested according to the ΔTm value.

[0149] 1. The main components of the hereditary deafness gene detection kit of the present invention are shown in Table 8, where the primers and probes are defined by the corres...