Multichannel fluorescent PCR detection kit for congenital deafness gene

A technology of congenital deafness and detection kits, applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., can solve problems that are not suitable for clinical application, prevent false negatives, avoid mutual interference, and reduce hydrolysis pressure Effect

Inactive Publication Date: 2015-06-10
JINAN YING SHENG BIOTECH
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  • Description
  • Claims
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Problems solved by technology

ARMS-PCR and RFLP methods require electrophoresis analysis...

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  • Multichannel fluorescent PCR detection kit for congenital deafness gene
  • Multichannel fluorescent PCR detection kit for congenital deafness gene
  • Multichannel fluorescent PCR detection kit for congenital deafness gene

Examples

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Embodiment 1

[0057] Example 1: Composition and detection method of a congenital deafness gene multi-channel fluorescent PCR detection kit

[0058] 1. The composition of the kit:

[0059] 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);

[0060] 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.

[0061] Wherein, the primers are primer 1, primer 2, primer 3 and internal reference primers, and their sequences are respectively:

[0062] Primer 1F: 5'-ATGGATTGGGGCACGCTG-3' (SEQ ID NO: 1),

[0063] Primer 1R: 5'-GACCTTCTGGGTTTTGAT-3' (SEQ ID NO: 2)

[0064] Primer 2F: 5'-GTACGACGGCTTCTCCAT-3' (SEQ ID NO: 3)

[0065] Primer 2R: 5'-GACAGTCTTTCTCCGTGGG-3' (SEQ ID NO: 4)

[0066] Primer 3F: 5'-CTCTTCCTCTACCTGCTGC-3' (SEQ ID NO: 5)

[0067...

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Abstract

The invention discloses a multichannel fluorescent PCR detection kit for a congenital deafness gene. With 35delG, 155delTCTG,176dell6bp, 235delC, 299-300delAT and 512insAACG mutations of a GJB2 gene, and 538G>T and 547G>A mutations of a GJB3 gene as detection objects, specific primers and a probe are designed, wherein the probe is used for specifically detecting whether 35, 155, 176, 235 and 299 loci of the GJB2 gene, and 538 and 547 loci of the GJB3 gene contain mutations or not. According to the multichannel fluorescent PCR detection kit, eight mutation loci of the congenital deafness are detected at the same timefor the first time in a same reaction tube by applying a multichannel fluorescent PCR method; the operation is simple, convenient, fast, accurate, high in flux, and low in cost; cross contamination is avoided by a pipe closing operation; and the deafness screening is high in intentionality and wide in screening range.

Description

technical field [0001] The invention relates to a congenital deafness gene multi-channel fluorescent PCR detection kit, which belongs to the technical field of gene detection. Background technique [0002] Deafness is one of the common clinical genetic diseases, which seriously affects the quality of human life. The second sample survey of disabled people in China in 2006 showed that the total number of people with disabilities in the country is as high as 80 million, and there are currently 27.8 million people with hearing and speech disabilities, accounting for about 35% of the total number of people with disabilities, including 20.04 million people with simple hearing disabilities. Among those with hearing and speech disabilities, there are 800,000 deaf children under the age of 7, and the number of deaf children is increasing at an annual rate of 50,000. It can be seen that the current situation of the hearing loss population in my country is particularly serious. [0...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q1/6827
Inventor 冯振景叶松弭兆元史桂芝
Owner JINAN YING SHENG BIOTECH
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