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Detection kit and detection method for metabolic marker of pegylated interferon alpha

A technology of metabolic markers and detection kits, applied in the field of gene detection, can solve the problems of cumbersome operation and time-consuming

Inactive Publication Date: 2021-12-07
菲思特(上海)生物科技有限公司
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

At present, the methods of obtaining blood DNA mainly include traditional column extraction and magnetic bead extraction, both of which are time-consuming and cumbersome to operate

Method used

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  • Detection kit and detection method for metabolic marker of pegylated interferon alpha
  • Detection kit and detection method for metabolic marker of pegylated interferon alpha
  • Detection kit and detection method for metabolic marker of pegylated interferon alpha

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Embodiment 2

[0059] Embodiment 2, kit detection steps

[0060] The instruments adopted in the present invention are as follows: thermostat, pyrosequencer (Wuhan First Biotechnology Co., Ltd.).

[0061] 1) Take 30 μL EDTA anticoagulated whole blood sample in a PCR amplification tube;

[0062] 2) Add 100 μL of sample treatment solution and 4ul of magnetic beads, and let stand for 5 minutes;

[0063] 3) Place the PCR amplification tube on the magnetic stand, and suck out the mixed solution from the opposite side of the magnetic beads after all the magnetic beads are adsorbed to one side;

[0064] 4) Add amplification reagent 1 into the dry powder of amplification reagent 2, fully dissolve and mix;

[0065] 5) Add 25ul of the prepared PCR reaction solution into the PCR amplification tube obtained in step 4), fully mix the magnetic beads and the PCR reaction solution, centrifuge, and perform constant temperature reaction.

[0066] 6) Amplify using a PCR instrument, the reaction system is 25 ...

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Abstract

The invention discloses a detection kit for metabolic markers of pegylated interferon alpha. A specific amplification primer and a sequencing primer are designed by the detection kit according to the gene polymorphism of the metabolic markers IFNL3rs12979860, IFNL3rs8099917 and IFNL3rs11881222 of the pegylated interferon alpha, and the gene polymorphism is detected through pyrosequencing. The kit comprises the following components: a sample treatment solution, magnetic beads, an amplification reagent 1, an amplification reagent 2, an IFNL3rs12979860 sequencing primer, an IFNL3rs8099917 sequencing primer, an IFNL3rs11881222 sequencing primer and a positive control. According to the detection kit and detection method for the metabolic markers of the pegylated interferon alpha, biotin labeled single-stranded DNA is combined with streptavidin, sequencing primers and sequencing raw materials are added after washing, pyrosequencing is carried out, the sequencing process is simplified, the sequencing time is shortened, the gene polymorphism for predicting the curative effect of the pegylated interferon alpha is detected by combining a rapid DNA preparation technology, a constant-temperature PCR amplification technology and a pyrosequencing technology. Suggestions from a gene perspective are provided for clinical personalized medication.

Description

technical field [0001] The invention relates to a detection kit for a peginterferon alpha metabolic marker and a detection method thereof, belonging to the field of gene detection. Background technique [0002] IL28B is located on chromosome 19, and its encoded product is interferon-λ3 (IFNL3), which is a member of the type III interferon family and has biological activities such as anti-virus, anti-tumor and immune regulation. There are SNP sites (rs12979860, rs8099917 and rs12980275) on the IL28B gene, which can affect gene expression, and then affect disease susceptibility and drug efficacy. IL28B belongs to the IFNXs family. IFNXs mainly induce the phosphorylation of STATI and STAT2 through the JAK-STAT signaling pathway, and promote myxovirus resistance protein A (myxovirus resistance protein A, MxA) and 2', 5'-oligoadenylate synthase The expression of the gene family, thereby exerting the antiviral effect two. Studies have confirmed that IFNλ3 can effectively inhibit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/686C12Q1/6869C12N15/11
CPCC12Q1/6883C12Q1/686C12Q1/6869C12Q2600/156C12Q2600/106C12Q2523/308C12Q2521/507C12Q2527/101C12Q2563/143C12Q2563/149C12Q2565/301
Inventor 孙悦陈立波刘丹
Owner 菲思特(上海)生物科技有限公司
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