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Detection method, kit and application of polymorphism in 1p36.22 region associated with hepatocellular carcinoma

A technology for detecting kits and polymorphic sites, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., to achieve the effect of reducing the risk of occurrence

Active Publication Date: 2011-12-14
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is no research report on the correlation between the genomic region 1p36.22 (UBE4B-KIF1B-PGD) polymorphism and the susceptibility to HBV-related HCC.

Method used

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  • Detection method, kit and application of polymorphism in 1p36.22 region associated with hepatocellular carcinoma
  • Detection method, kit and application of polymorphism in 1p36.22 region associated with hepatocellular carcinoma
  • Detection method, kit and application of polymorphism in 1p36.22 region associated with hepatocellular carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] In this example, a set of amplification primer pairs and an extension primer were designed and synthesized, and based on these primers, a method for detecting the susceptibility genomic region 1p36.22 (UBE4B-KIF1B-PGD) associated with the risk of HCC was designed ) method of polymorphic site rs17401966 genotype.

[0047] 1. Genomic DNA extraction

[0048] Genomic DNA of peripheral blood leukocytes was extracted by the phenol / chloroform method in the field.

[0049] 1) Take sodium citrate or EDTA-Na 2 5ml of anticoagulated whole blood (try not to use heparin for anticoagulation), put in a 50ml capped centrifuge tube;

[0050] 2) Add 3-5 times the volume of cold distilled water, invert and mix repeatedly, place in ice for 5 minutes, and centrifuge at 2000rpm at 4°C for 20 minutes;

[0051] 3) Slowly pour the supernatant, add pre-cooled 0.1% Triton-X100 (same volume as above) to the precipitate, gently mix the precipitate, centrifuge as above, and discard the supernatan...

Embodiment 2

[0091] In this example, a set of amplification primer pairs and a pair of probes were designed and synthesized, and on the basis of these primers and probes, a method for detecting the susceptibility genomic region 1p36.22 (UBE4B) associated with the risk of HCC was designed. -KIF1B-PGD) polymorphic site rs17401966 genotype method.

[0092] 1. Genomic DNA extraction

[0093] With embodiment 1.

[0094] 2. Primer and probe synthesis

[0095] Design the primer pair and probe sequence according to the genomic region where rs17401966 is located, among which the forward primer:

[0096] 5'-TTTCCAGCACTTAATGAAAACACATAG-3', reverse primer:

[0097] 5'-CAAAGTTAAATTTCCCTGCTTTGAA-3', MGB probe 1:

[0098] 5'(FAM)-TATGAGTCCATATTGAGTC-3', MGB probe 2:

[0099] 5'(HEX)-TATGAGTCCGTATTGAGT-3'. All primers and probes were synthesized by commercial companies (such as Shanghai Jikang Co., Ltd., etc.), using ddH 2 O Dilute the primers to 20 μM and the probe to 10 μM. Note: Probes should b...

Embodiment 3

[0111] In this example, the methods established in Examples 1 and 2 are used to analyze the rs17401966 genotypes of six populations.

[0112] 1. Test object

[0113] Five independent case-control populations (a total of 2,317 cases and 1,790 controls) and one core family population (159 core families) were included in the study. All participants were Chinese adults, and the Guangxi case-control population, Guangxi pedigree, and Guangdong case-control population were recruited from southern China, the Shanghai case-control population and Jiangsu case-control population were recruited from eastern China, and the Beijing case-control population was recruited from northern China. All HCC cases were chronic HBV carriers, newly diagnosed, not treated with radiotherapy and chemotherapy, and not combined with other tumors. The diagnostic criteria for HCC include: positive tissue biopsy; serum alpha-fetoprotein level ≥ 400 ng / ml, combined with a positive imaging diagnosis. The contro...

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Abstract

The invention relates to a detection method, kit and application thereof for the polymorphic site rs17401966 in the susceptibility genome region 1p36.22 related to the risk of HBV-related hepatocellular carcinoma (Hepatocellular carcinoma, HCC). The method is SNPstream typing and TaqMan typing, and the present invention further relates to a kit for detecting the above-mentioned susceptible sites and its application. In addition, the present invention associates the polymorphic site rs17401966 in the 1p36.22 region with the risk of HBV-related HCC, and determines that 1p36.22 is a new susceptibility gene region of HBV-related HCC, which is the key factor for HBV-related HCC. Population prevention and control provide new genetic counseling content.

Description

technical field [0001] The present invention relates to individual susceptibility in population prevention of hepatocellular carcinoma (Hepatocellular carcinoma, HCC) and its genetic detection method, kit and application thereof. Specifically, the present invention relates to a method for detecting the rs17401966 genotype of the susceptibility genomic region 1p36.22 (UBE4B-KIF1B-PGD) associated with the risk of HBV-related HCC; the present invention further relates to a detection reagent for the genotype of the above susceptibility genomic region In addition, the present invention also relates to the application of the above susceptibility genomic regions, related detection methods and detection kits in the prevention and control of HCC and genetic counseling. Background technique [0002] HCC is one of the most common cancers and the fifth leading cause of cancer death worldwide. HCC is rarely diagnosed early and usually dies within months of diagnosis. It is estimated th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 周钢桥张红星贺福初翟芸
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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