Primer combination used for guiding lovastatin drug individualized medication related gene detection and kit and method

A lovastatin and gene detection technology, which is applied in the field of primer combinations and kits for guiding lovastatin individualized drug-related gene detection, can solve the problems of no kit products, adverse reactions of clinical medication, etc., and reduce medical costs , detection of fast, rapid and accurate determination of the effect

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A lovastatin and gene detection technology, which is applied in the field of primer combinations and kits for guiding lovastatin individualized drug-related gene detection, can solve the problems of no kit products, adverse reactions of clinical medication, etc., and reduce medical costs , detection of fast, rapid and accurate determination of the effect

CN110295225AInactive Publication Date: 2019-10-01南京派森诺基因科技有限公司

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Experimental program

Comparison scheme

Effect test

Embodiment 1

[0081] Embodiment 1: the preparation of kit

[0082] 1. Design and synthesis of primers and probes

[0083] For the APOA5 (rs662799), CETP (rs708272), SLCO1B1 (rs2291073) gene loci in the human genome (for the sequence, refer to the human genome sequence published in the NCBI database), use Primer Premier 5.0 and PrimerExpress 3.0 software to design specific primers and Taqman probe.

[0084] Specific primers and probe sequences are shown in the following table 1:

[0085] Table 1 Primer and probe sequence and length information

[0086]

[0087]

[0088] 2. Selection of reference substance

[0089] The positive control substance one is respectively APOA5 (rs662799), CETP (rs708272), and SLCO1B1 (rs2291073) gene site wild-type plasmids;

[0090] The two positive control substances are respectively APOA5 (rs662799), CETP (rs708272), and SLCO1B1 (rs2291073) plasmids heterozygous for the wild type and mutant type at a quantitative ratio of 1:1;

[0091] The three posit...

Embodiment 2

[0104] Example 2: Using the above kit for lovastatin drug individualized drug-related gene detection method

[0105] 1. Biological samples

[0106] The biological materials used in the present invention are all from Parsonuo Medical Laboratory. Anticoagulant blood was used for the remaining 500 cases tested at the Paisino Medical Laboratory during September-December 2018.

[0107] 2. Take the DNA extracted from the sample to be tested as a PCR template:

[0108] The DNA extraction kit is the extraction kit "Blood Genomic DNA Extraction Kit" produced by Tiangen Biochemical Technology (Beijing) Co., Ltd. (Cat. No.: DP318).

[0109] 1) Take 400ul of whole blood, add 800ul of cell lysate CL, mix well, centrifuge at 10000rpm / 11500×g for 1min, discard the supernatant, and repeat once if the lysis is not complete.

[0110] 2) Add 200ul buffer GS to the precipitation, mix well, then add 20ul proteinase K, mix well.

[0111] 3) Add 200ul buffer solution GB, mix well, 56°C, 10min, i...

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Abstract

The invention discloses a primer combination used for guiding lovastatin drug individualized medication related gene detection. The primer combination includes specific primers for APOA5(rs662799), CETP(rs708272) and SLCO1B1(rs2291073) gene loci and Taqman probes. The invention further discloses a kit comprising the primer combination and an analysis method thereof. The primer combination has theadvantages of being easy to operate, economical, rapid in detection, high in accuracy, good in specificity and simple in result interpretation, rapid and accurate determination for the lovastatin drugindividualized medication related gene polymorphism is achieved, so that adverse reactions of clinical lovastatin drug common medication are reduced, the medical cost is lowered, and the social resources are saved.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a primer combination, a kit and a method for guiding lovastatin drug individualized drug-related gene detection. Background technique [0002] Hypercholesterolemia refers to blood cholesterol levels exceeding the normal value (5.7mmol / L). Long-term high cholesterol will cause atherosclerosis in the body, which will cause coronary heart disease, cerebral infarction and other diseases that will endanger life. [0003] Hyperlipidemia mainly refers to elevated blood cholesterol and / or triglycerides. Hyperlipidemia is a manifestation of abnormal fat metabolism in the human body, and is mainly divided into three categories: hypercholesterolemia, hypertriglyceridemia, and mixed hyperlipidemia. Hyperlipidemia is a common metabolic disease in clinical practice. It is a disease in which one or more lipids in plasma are higher than the normal index due to abnormal fat metabolis...

Claims

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Application Information

Patent Timeline

01 Oct 2019

Publication

CN110295225A

IPC: C12Q1/6883; C12Q1/6858; C12N15/11

CPC: C12Q1/6883; C12Q1/6858; C12Q2600/106; C12Q2600/156; C12Q2600/166; C12Q2531/113; C12Q2563/107; C12Q2545/113

Inventors: 刘颖; 杜晴晴