Drug-induced deafness gene multichannel fluorescence PCR (polymerase chain reaction) detection kit

A technology for drug-induced deafness and detection kits, applied in the field of genetic testing, can solve problems such as being unsuitable for clinical application, and achieve the effect of preventing false negatives

Inactive Publication Date: 2015-06-17
JINAN YING SHENG BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ARMS-PCR and RFLP methods require electrophoresis analysis of PCR products, which are not suitable for clinical application

Method used

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  • Drug-induced deafness gene multichannel fluorescence PCR (polymerase chain reaction) detection kit
  • Drug-induced deafness gene multichannel fluorescence PCR (polymerase chain reaction) detection kit
  • Drug-induced deafness gene multichannel fluorescence PCR (polymerase chain reaction) detection kit

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Embodiment 1

[0051] Example 1: Composition and detection method of a drug-induced deafness gene multi-channel fluorescent PCR detection kit

[0052] 1. The composition of the kit:

[0053] 1 tube of DNA extraction solution (5.0ml), 1 tube of PCR reaction solution (1.1ml), 1 tube of negative quality control (50μl), 1 tube of positive quality control (50μl);

[0054] The composition of each 50 μl PCR reaction solution is: 5.0 μl of 10×PCR buffer, MgCl 2 3.0mM, dNTPs 1.0mM, primer 1.0μM, probe 0.5μM, Taq enzyme 3.0U, UNG enzyme 1.0U, add water to 50μl.

[0055] Wherein, the primers include primer 1, primer 2 and internal reference primers, the sequences of which are respectively:

[0056] Primer 1F: 5'-GGCCCTGAAGCGCGTACA-3' (SEQ ID NO: 1)

[0057] Primer 1R: 5'-AAGTGTAAGTTGGGTGCT-3' (SEQ ID NO: 2)

[0058] Primer 2F: 5'-ATCAAGCACGCAGCAATG-3' (SEQ ID NO:3)

[0059] Primer 2R: 5'-GTGTTCTGGCGAGCAGTTT-3' (SEQ ID NO: 4)

[0060] β-ActinF:5'-CACCCAGCACAATGAAGA-3' (SEQ ID NO:9)

[0061] β-Act...

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Abstract

The invention discloses a drug-induced deafness gene multichannel fluorescence PCR (polymerase chain reaction) detection kit. Two pairs of specific primers and four TaqMan probes are designed according to four sites on the drug-induced deafness related gene mtDNA 12S rRNA: 1555A>G, 1494C>T, 1095T>C and 827A>G, and a pair of specific primers and probes are designed by using the housekeeping gene beta-actin as the internal reference to monitor the effectiveness of the sample template and reaction system and avoid the appearance of false negative; and meanwhile, the setting of the negative and positive quality control ensures the accuracy of the detection result. The kit implements simultaneous detection on four mutant sites of the drug-induced deafness gene in one reaction tube by a multichannel fluorescence PCR process for the first time. The method is simple to operate, has the advantages of high speed, high accuracy, high flux and low cost, avoids the cross contamination due to shut tube operation, improves the purposiveness and screening range of deafness screening, effectively prevents the drug-induced deafness, and is easy for popularization and application.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a kit for gene detection of drug-induced deafness based on a multi-channel fluorescent PCR method. 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] Individuals carrying the mutation gene for drug-induced deafness are sensitive to aminoglycoside antibiotics, and the application of such drugs can lead to substantial hearing loss, and even the clinically common "one-shot deafness" phenomenon. [0004] Aminoglycoside-related...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 冯振景叶松史桂芝弭兆元
Owner JINAN YING SHENG BIOTECH
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