A multi-channel fluorescent PCR detection kit for delayed deafness gene

Abstract

The invention discloses a multichannel fluorescent PCR detection kit of a delayed deafness gene. Aiming at 7 sites on the delayed deafness gene PDS, namely IVS7-2A>G, 2168A>G, 1174A>T, 1229C>T, 1226G>A, 1975G>C and 2027T>A, 4 pairs of specific primers and 7 TaqMan probes are designed, with the human housekeeping gene beta-Actin as reference, 1 pair of specific primer and probe is designed, and the accuracy of a detection result is guaranteed by monitoring the effectiveness of a sample template and a reaction system, avoiding false negative and setting negative and positive quality control at the same time. The multichannel fluorescent PCR method is used for detecting 7 mutation sites of delayed deafness in the same reaction tube for the first time, the method is operated quickly, conveniently and accurately, and is high in throughput and low in cost, the operation is carried out in a closed tube to avoid cross contamination, so that the deafness screening teleonomy is strong and the screening scope is wide, and thus the delayed deafness is effectively prevented, and the multichannel fluorescent PCR detection kit is easy to popularize and apply.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a delayed-onset deafness gene multi-channel fluorescent PCR detection kit. Background technique [0002] Deafness is one of the common clinical disability diseases. Clinically, deafness can be divided into three types according to the nature, cause and lesion site affected by the auditory system: conductive deafness (outer ear or middle ear lesions), sensorineural deafness (inner ear lesions), mixed deafness. Research data show that sensorineural deafness and mixed deafness are closely related to genetics, and deafness caused by genetics accounts for more than 60% of deafness. [0003] Patients with delayed deafness carry the PDS gene, and may develop severe deafness or total deafness under the stimulation of predisposing factors (head trauma, strenuous exercise, cold, etc.) during their growth. The genetic mode of delayed deafness is autosomal recessive inheritance, and the gene ...

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Problems solved by technology

[0006] Chinese patent "Method and kit for detection of IVS7-2A>G mutation of PDS gene by fluorescence PCR technology" (CN104120167A) discloses a method and kit for detection of deafness gene PDS gene IVS7-2A>G mutation by fluorescence quantitative PCR technology , but one PCR reaction tube of this kit can only detect one mutation site on the deafness gene PDS, and cannot detect other sites such as 2168A>G, 1229C>T, 1174A>T with a higher mutation rate, thus reducing the delay Screening Coverage for Acute Deafness

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