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Probe for detecting mutation of gene ALDH2, as well as application thereof and kit

A technology for detecting probes and kits, which is applied in the field of probes for detecting ALDH2 gene mutations and their applications and kits, can solve the problems of contamination, inability to determine the location and type of SNP, expensive special instruments, etc., and achieves simple and convenient operation. The effect of low cost and low false positive rate

Inactive Publication Date: 2017-05-10
GUANGZHOU EZLIFE BIO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it can only determine whether there is a mutation, and cannot determine the position and type of SNP. It needs to be verified by standard samples or combined with sequencing, and requires expensive special instruments and professional analysis software. This is one of the reasons why DHPLC has not been widely implemented.
In addition, the detection process needs to open the reaction tube, which is easy to cause pollution

Method used

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  • Probe for detecting mutation of gene ALDH2, as well as application thereof and kit
  • Probe for detecting mutation of gene ALDH2, as well as application thereof and kit
  • Probe for detecting mutation of gene ALDH2, as well as application thereof and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] The probes for detecting ALDH2 gene mutations provided in this example include wild-type capture probes and detection probes, and the 5' end of the detection probes is labeled with biotin (Biotin).

[0061] The base sequence of the wild-type capture probe is as follows:

[0062] 5'-ATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT TCACTTCAGTGTA -3' (SEQ ID NO.2), wherein the underlined part is used for the target region (5'-TACACT) of the target sequence (SEQ ID NO.4) covering the rs671 site (G) of the wild-type ALDH2 gene G AAGTGA-3') reverse complementary binding, and then capture and immobilize the wild-type target sequence.

[0063] In addition, the 1-77th thymine (T)-rich region at the 5' end of the wild-type capture probe is an extended arm, which is used to increase the probability of capture and binding to the wild-type target sequence, thereby improving the sensitivity of detection.

[0064] The base sequence of the wild-type targe...

Embodiment 2

[0070] The probes for detecting ALDH2 gene mutations provided in this example include mutant capture probes (SEQ ID NO.3) and detection probes (SEQ ID NO.1), and the base sequence and structure of the detection probes are the same as in Example 1 .

[0071] The base sequence of the mutant capture probe is as follows:

[0072] 5'-ATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT TCACTTTAGTGTA -3' (SEQ ID NO.3), wherein, the underline part is used for the target region (5'- TACACT A AAGTGA-3', the underline is the rs671 site (A)) reverse complementary binding, and then capture and fix the mutant target sequence.

[0073] In addition, the 1-77th thymine (T)-rich region at the 5' end of the mutant capture probe is an extension arm, which can be used to improve the interaction between the mutant capture probe and the mutant target sequence (SEQ ID NO.5) capture binding probability, improve detection sensitivity.

[0074] The base sequence of the sel...

Embodiment 3

[0078] The probes for detecting ALDH2 gene mutations provided in this example include: wild-type capture probes, mutant capture probes and detection probes.

[0079] The base sequence of the wild-type capture probe is the same as in Example 1, the base sequence of the mutant capture probe is the same as in Example 2, and the base sequence and structure of the detection probe are the same as in Example 1.

[0080] The probe provided in this embodiment can not only detect the mutation of the ALDH2 gene rs671 site of the sample to be tested, but also can distinguish whether the sample to be tested is a wild type homozygote, a mutant heterozygote or a mutant homozygote, It also has the characteristics of good specificity, high sensitivity and low false positive rate.

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Abstract

The invention discloses a probe for detecting mutation of a gene ALDH2, as well as application thereof and a kit, and belongs to the technical field of gene detection. The probe comprises a detection probe as shown by SEQ ID NO.1, and one or two of capture probes as shown by SEQ ID NO.2-3. The probe can detect G to A mutation at site rs671 of the gene ALDH2, and has the characteristics of high detection sensitivity, strong specificity, low false positive rate, and the like.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a probe for detecting ALDH2 gene mutation, its application and a kit. Background technique [0002] Acetaldehyde dehydrogenase (Acetaldehyde Dehydrogenase, ALDH) is encoded by the ALDH gene and can be divided into ALDH1A1, ALDH1B1 and ALDH2. Among them, ALDH2 has the strongest ability to catalyze acetaldehyde into acetic acid in the human body. The ALDH2 gene is located on chromosome 12 (12q24), consists of 13 exons, and encodes an enzyme protein peptide chain consisting of 500 amino acid residues. Studies have shown that polymorphisms in this gene are common in Asian populations and may affect drinking behavior in the population. ALDH2 is most closely related to the Asian population because of the G→A mutation at the rs671 site of the ALDH2 gene, which can cause the encoded polypeptide chain glutamic acid (Glu) to be lysine (Lys). The ALDH2 gene is related to the catab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6834C12Q1/6883C12Q1/6888C12Q2600/106C12Q2600/156C12Q2565/519C12Q2565/607
Inventor 廖玮莫亚勤林晓燕张晨光
Owner GUANGZHOU EZLIFE BIO CO LTD
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