Primers, kit and method for detection of polymorphism of human IL28B gene

A gene polymorphism and detection method technology, applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of complicated operations, prone to deviations in results, and long time-consuming, etc., to achieve suitable Clinical promotion, improvement of diagnosis and treatment level, and high clinical application value

Inactive Publication Date: 2015-07-15
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Abstract
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  • Application Information

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Problems solved by technology

The above-mentioned disclosed detection methods all use conventional methods such as gene sequencing, which are complicated to o

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  • Primers, kit and method for detection of polymorphism of human IL28B gene
  • Primers, kit and method for detection of polymorphism of human IL28B gene
  • Primers, kit and method for detection of polymorphism of human IL28B gene

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Embodiment 1

[0053] Embodiment 1 According to the assembly of human IL28B gene polymorphism detection kit of the present invention

[0054] Generally, the human IL28B gene polymorphism detection kit according to the present invention may include the following general reagents and consumables:

[0055]

[0056] Specifically, the main reagents in the human IL28B gene polymorphism detection kit according to the present invention include:

[0057] (1) PCR amplification reagents: 2×PCR mix, HotstarTaq enzyme, pure water and PCR amplification primers for detecting polymorphisms of rs12979860 and rs8099917 sites, wherein,

[0058] The rs12979860PCR upstream primer sequence is:

[0059] 5'-ACGTTGGATGTCGTGCCTGTCGTGTACTGA-3' (SEQ ID NO: 1);

[0060] The downstream primer sequence of rs12979860PCR is:

[0061] 5'-ACGTTGGATGAGCGCGGAGTGCAATTCAAC-3' (SEQ ID NO: 2).

[0062] The rs8099917 PCR upstream primer sequence is:

[0063] 5'-ACGTTGGATGCAATTTGTCACTGTTCCTCC-3' (SEQ ID NO: 4);

[0064] The ...

Embodiment 2

[0070] Embodiment 2: The detection example of human IL28B gene polymorphism detection kit according to the present invention

[0071] test sample : EDTA anticoagulated blood samples were collected from healthy adults with routine physical examinations from the Physical Examination Department of Shenzhen Second People's Hospital.

[0072] Main equipment :

[0073] 1. MassARRAY Genetic Analysis System (SEQUENOM Inc., USA);

[0074] 2. AB-Vii A7 fluorescent quantitative PCR instrument (Applied Biosystems (ABI));

[0075] 3. Centrifuge (TL-5.0W, Shanghai);

[0076] 4. Rotary oscillator (Shanghai).

[0077] Detection procedure :

[0078] 1. Purification of genomic DNA from blood samples: extract using Tiangen Biochemical Blood Genome Extraction Kit.

[0079] 2. PCR amplification: use the purified blood sample DNA as a template, and perform PCR amplification with the PCR amplification primer pairs respectively targeting the polymorphisms of rs12979860 and rs8099917 sites ...

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Abstract

The invention relates to PCR amplimers, single base extension primers, a detection kit and a detection method for the polymorphism of the human IL28B gene. The kit comprises the PCR amplimers and the single base extension primer designed specific to the polymorphism of the site rs12979860 (T/C), and the PCR amplimers and the single base extension primer designed specific to the polymorphism of the site rs8099917 (T/G). The above-mentioned primers are used for PCR amplification, SAP reaction and single base extension, and then mass spectrometry is employed for analysis of the forms of the polymorphism of the above-mentioned sites. The detection method for the polymorphism of the human IL28B gene can rapidly, specifically and highly efficiently detect the polymorphism of the two above-mentioned SNP sites of the IL28 gene; thus, prediction can be carried out on the curative effects of a standard hepatitis C therapy scheme, differentiated dealing and treatment can be carried out on patients with different types of hepatitis C, and the whole diagnosis and treatment level of hepatitis C is improved.

Description

technical field [0001] The invention belongs to the technical field of biomedical molecular detection, and in particular relates to a detection primer, a kit and a detection method for the detection of nucleotide polymorphisms at two sites of rs12979860 and rs8099917 in the human IL28B gene. Background technique [0002] Viral hepatitis C is an infectious disease caused by hepatitis C virus (HCV), which is mainly transmitted through intravenous drug injection, blood transfusion, organ transplantation and hemodialysis. At present, there are about 130 to 170 million hepatitis C patients in the world, and as many as 350,000 people die from HCV infection every year. Compared with hepatitis B, hepatitis C is more likely to become chronic. It has become one of the main causes of liver cirrhosis and liver cancer, and seriously endangers human health. Therefore, active prevention and effective treatment of hepatitis C are very necessary. [0003] The specific mechanism of HCV infec...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6827C12Q1/6888C12Q2600/106C12Q2600/156
Inventor 许蕴刘文兰黄建林李赟
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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