Composition for detecting gene polymorphism of SLCO1B1 and ApoE as well as application thereof

A technology of SLCO1B1388-AFP and SLCO1B1521, which is applied in the field of detecting SLCO1B1 and ApoE gene polymorphisms, can solve the problems of adding probes, low accuracy and specificity, and achieve the effects of avoiding cross-contamination, accurate results, and improving simplicity

Active Publication Date: 2018-01-05
北京鑫诺美迪基因检测技术有限公司
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  • Abstract
  • Description
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Problems solved by technology

And on this basis, the probe is modified by locking nucleic acid and secondary structure, increasing the Tm value of the probe, and further improving the specificity of the probe binding. Finally, through the adjustment of the above system, the accuracy and specificity are evenly balanced. It can reach 100%, and the minimum detection concentration can reach 1ng / μL, which solves the problem of low accuracy and specificity in the detection of SLCO1B1 and ApoE gene polymorphisms in the past

Method used

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  • Composition for detecting gene polymorphism of SLCO1B1 and ApoE as well as application thereof
  • Composition for detecting gene polymorphism of SLCO1B1 and ApoE as well as application thereof
  • Composition for detecting gene polymorphism of SLCO1B1 and ApoE as well as application thereof

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Embodiment 1

[0113] Embodiment 1: On the basis of multiplex fluorescent PCR, the present invention is formulated into a premixed primer composition for detection sites. The extracted sample can be tested directly, which makes the detection step easier.

[0114] Specific steps are as follows:

[0115] 1) Genomic DNA was extracted from the whole blood sample (using a commercial peripheral blood genomic DNA extraction kit).

[0116] 2) Measure the concentration of the extracted genomic DNA, and dilute the concentration to 10-20 ng / μL for subsequent PCR reaction process.

[0117] 3) Composition of the reaction system: dNTPs, Mg 2+ , DNA polymerase buffer, reverse transcription buffer, gene primer pair, DNA polymerase, etc.

[0118] 4) PCR reaction conditions: UDG enzyme reaction at 37°C for 2min; pre-denaturation at 95°C for 3min; denaturation at 94°C for 15s, annealing and extension at 60°C for 35s, 40 cycles.

[0119] 5) Preparation of PCR reaction system:

[0120] PCR amplification MIX...

specific Embodiment approach 2

[0145] Collect 200 peripheral blood control samples from Wuhan Tongji Hospital, and use the above-mentioned finished kit for detection. The specific operation steps are as described in Example 1.

[0146] detection gene

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Abstract

The invention discloses a composition for detecting gene polymorphism of SLCO1B1 and ApoE as well as an application thereof. An FAM, HEX, ROX (modified) three-channel multiple PCR reaction is used fordetecting single nucleotide polymorphism sites in SLCO1B1 and ApoE genes. In order to improve simplicity and specificity of detection, a probe typing technological means is used for realize one-tubetyping detection, and the whole operation and reaction process are simplified; on the basis, locked nucleic acid modification and secondary structure modification are carried out for the probe, Tm value of the product is added, in order to further improve specificity of probe combination; and finally proportioning of primer pairs, amount of Taq enzyme, amount of magnesium ions and the like are adjusted, in order to improve lowest detection limit, accuracy and specificity of the whole kit, lowest detection concentration of the kit reaches 1ng / [mu]L, and accuracy and specificity can reach 100%.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a technology for detecting SLCO1B1 and ApoE gene polymorphisms. Background technique [0002] Statins are hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, which block the endogenous cholesterol synthesis rate-limiting enzyme (HMG-CoA) reductase by competitively inhibiting The metabolic pathway of intracellular valonate reduces intracellular cholesterol synthesis, thereby stimulating the increase in the number and activity of low-density lipoprotein (LDL) receptors on the surface of cell membranes (mainly liver cells) and increasing serum cholesterol clearance. level lowered. Statins can also inhibit the synthesis of apolipoprotein B-100 in the liver, thereby reducing the synthesis and secretion of triglyceride-rich AV and lipoproteins. [0003] Statins are divided into natural compounds (such as lovastatin, simvastatin, pravastatin, mevastatin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 刘沛王林海
Owner 北京鑫诺美迪基因检测技术有限公司
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