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A human chromosome gene il28b site polymorphism detection kit and its detection method and application

A chromosomal gene and locus polymorphism technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. Uncertainty of test results and other problems, to shorten the test time, reduce production costs, and avoid inaccurate test results

Inactive Publication Date: 2016-06-01
厦门安普利生物工程有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The above existing technologies all have uncertainty in the detection results, and at the same time, they can only detect polymorphic sites with insufficient information, and the detection process is complicated and has limitations.

Method used

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  • A human chromosome gene il28b site polymorphism detection kit and its detection method and application
  • A human chromosome gene il28b site polymorphism detection kit and its detection method and application
  • A human chromosome gene il28b site polymorphism detection kit and its detection method and application

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Embodiment 1

[0077] 1. Lysis: Take 100 μl of ethylenediaminetetraacetic acid (EDTA) tripotassium salt or sodium citrate anticoagulated whole blood from the patient to be tested (Xiamen Hospital of Traditional Chinese Medicine) and add it to a 1.5ml centrifuge tube, then add 200 μl of lysis buffer (3M Melonium isothiocyanate, 10mM EDTA, 1% Triton-X100, 20mM Tris-HCl (pH 6), 20μl proteinase K, mixed evenly, heated at 55°C for 10min. 2. Magnetic adsorption, add 10 μl magnetic nanoparticles (Shanghai Aorun Micro-Nano New Material Technology Co., Ltd. SM-015D) to the above centrifuge tube, mix well and let it stand for 10 minutes, place it on a magnetic stand for magnetic separation, and absorb the waste liquid , suck as much as possible. 3. Washing, add 300 μl washing liquid (1.5MNaCl, 0.5% Triton-X100, 10mM EDTA, 20mM Tris-HClpH6) to the above-mentioned centrifuge tube, shake and mix, put it on a magnetic adsorption rack, solidify the magnetic particles, and absorb the waste liquid. 4. For e...

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Abstract

The invention provides a human chromosome gene IL28B site polymorphism detection kit, and a detection method and application thereof. The kit comprises at least one group of primers and probes in group a, group b and group c. The primers and the probes comprise: a: 860L1, 860L2, 860R, 860P; b: 275L1, 275L2, 275R, 275P; and c: 917L1, 917L2, 917R, 917P. 5' terminals of 860P, 275P and 917P are labeled as FAM, and 3' terminals are labeled as DACYL. The invention also discloses the detection method and application of the kit. The method is relatively simple and rapid, and is easy for realizing whole-course automation; secondary pollution of PCR products is avoided, and pollution is reduced; the cost is relatively low and relatively economic; and present popularized equipment in the market can be utilized, and no new investment is needed.

Description

technical field [0001] The invention relates to an in vitro diagnostic reagent, in particular to a detection kit for the polymorphism of the human chromosome gene IL28B site, a detection method and an application thereof. Background technique [0002] The polymorphisms of the host IL28B gene, and its closely related single nucleotide polymorphisms mainly include rs12979860, rs12980275, and rs8099917 located upstream of the IL28B gene on chromosome (19q13). [0003] At present, the detection methods of IL28B polymorphism mainly include gene sequencing method (such as 201210234084.6 kit for detecting IL28BSNP12980275 polymorphism in patients with hepatitis C) and reverse hybridization method (such as 201010019477.5 Hepatitis C virus genotyping and IL28 polymorphism). Both of these methods need to amplify the target gene first, and then detect the amplified product a second time, so the operation is complicated. At the same time, the gene amplification product is more than 10...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q2531/113C12Q2535/137C12Q2563/107
Inventor 魏超魏劭
Owner 厦门安普利生物工程有限公司
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