A kind of human mthfr and mtrr gene polymorphism detection primer, probe, kit and method

A technology of gene polymorphism and detection kit, which is applied in biochemical equipment and methods, microbe measurement/testing, DNA/RNA fragments, etc., can solve the problem of high instrument requirements, unfavorable large-scale promotion and cumbersome operation of gene chip method and other problems to achieve the effects of rapid detection, avoidance of cross-contamination of samples, and high sensitivity

Active Publication Date: 2018-03-09
广州达晖生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the sequencing method is the gold standard for the detection of genetic polymorphisms, the operation is cumbersome, the workload is heavy, and the instruments are expensive, which is not conducive to popularization
The gene chip method requires high equipment and high cost, which is not conducive to large-scale promotion

Method used

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  • A kind of human mthfr and mtrr gene polymorphism detection primer, probe, kit and method
  • A kind of human mthfr and mtrr gene polymorphism detection primer, probe, kit and method
  • A kind of human mthfr and mtrr gene polymorphism detection primer, probe, kit and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Human MTHFR and MTRR gene polymorphism detection kit

[0048] The kit described in this embodiment includes the following components:

[0049] Public primers and probes: public primers UP1 (nucleotide sequence shown in SEQ ID NO.1) and UP2 (nucleotide sequence shown in SEQ ID NO.2), beta globin specific upstream primer globinF (SEQ ID NO.3) (Nucleotide sequence shown), beta globin specific downstream primer globinR (nucleotide sequence shown in SEQ ID NO.4), beta globin specific probe globin probe (nucleotide sequence shown in SEQ ID NO.5) ;

[0050] Specific primers and probes:

[0051] For MTHFR677C> MTHFR677 specific upstream primer MTHFR677F (nucleotide sequence shown in SEQ ID NO.6), MTHFR677 downstream primer MTHFR677R (nucleotide sequence shown in SEQ ID NO.7), MTHFR677C specific probe for qualitative detection of T gene polymorphism Needle MTHFR677Cprobe (nucleotide sequence shown in SEQ ID NO. 8), MTHFR677T specific probe MTHFR677Tprobe (nucleotide sequence...

Embodiment 2

[0062] Example 2 Method for detecting human MTHFR and MTRR gene polymorphism

[0063] Use the kit of Example 1 for detection, including the following steps:

[0064] 1. DNA preparation

[0065] Thirty people were randomly selected to draw peripheral blood and DNA with informed consent in accordance with medical routines. The kit of the present invention has no specified requirements for the extraction method of human genomic DNA. Generally, the conventional laboratory method (phenol-chloroform extraction method) or the kit can be used to extract human genomic DNA. Take appropriate amount of EDTA-K2 or sodium citrate anticoagulated human peripheral whole blood and extract human genomic DNA. The extracted DNA samples must meet the following requirements.

[0066] 1.1 The sample source of the kit of the present invention is human peripheral whole blood genomic DNA, the DNA concentration is 10-50ng / μL, and the purity A260 / A280 is between 1.8-2.0.

[0067] 1.2 Sample storage: DNA samples ...

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Abstract

The invention discloses a human MTHFR and MTRR gene polymorphism detection primer, probe, test kit and method and belongs to the technical field of in-vitro nucleic acid detection. The two most common single nucleotide polymorphisms 677C>T and 1298A>C of the MTHFR and the common mutation site 66A>G of the MTHFR are designed for qualitative detection, and a specific primer containing reference genes beta globin and a probe are designed. The test kit includes a detection primer and probe combination, a reference substance, a PCR reaction liquid and the like. Three independent multiplex PCR reactions are conducted for qualitative detection for the three gene polymorphism sites, namely, MTHFR677C>T, MTHFR1298A>C, and MTHFR 66A>G respectively. The detection primer, the probe, the test kit and the method are strong in specificity, high in sensitivity, simple, rapid and convenient for large-scale application and popularization.

Description

Technical field [0001] The invention belongs to the technical field of in vitro nucleic acid detection, and is specifically a primer, probe, kit and method for detecting human MTHFR and MTRR gene polymorphism. Background technique [0002] Folic acid belongs to the B vitamins. It is an essential substance for cell growth and reproduction and participates in many important metabolic reactions of the body. Once the body has folate deficiency or folate metabolism enzyme deficiency, it will cause abnormal cell cycle, abnormal DNA and protein methylation reaction, abnormal synthesis of DNA bases and other abnormal biochemical reactions, which directly or indirectly leads to new birth Neural tube defects in children or fetuses and cardiovascular diseases in adults. At present, the genes that have been proven to be closely related to folate metabolism are 5,10-methylenetetrahydrofolate reductase (MTHFR) and methionine synthase reductase (MTRR). 5,10-Methylenetetrahydrofolate reductase...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q2537/143C12Q2545/101C12Q2563/107
Inventor 李晓辉徐素茹杨祖婷
Owner 广州达晖生物技术股份有限公司
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