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Human HLA-B27 gene typing kit

A technology for HLA-B27 and genotyping, which is applied in the field of human HLA-B27 genotyping kits, can solve the problems of many steps in DNA extraction from whole blood, increased detection time and factors affecting the results, and time-consuming problems. Reduce the risk of contamination, reduce the time of experimental operation, and the effect of accurate and reliable results

Active Publication Date: 2014-10-22
JIANGSU WEIHE BIOTECH
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  • Claims
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AI Technical Summary

Problems solved by technology

The PCR-SSP method requires simple instruments, easy to carry out projects, easy to interpret results, and good accuracy, but the whole blood DNA extraction has many steps, takes a long time, and increases the detection time and factors that affect the results

Method used

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  • Human HLA-B27 gene typing kit
  • Human HLA-B27 gene typing kit
  • Human HLA-B27 gene typing kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1 Raw materials and equipment:

[0031] 1.1 Contents of the kit:

[0032] 1) 8-well 200ul PCR reaction plate;

[0033] 2) 13 pairs of specific upstream and downstream amplification primers are freeze-dried at the bottom of the PCR reaction plate in a specific arrangement, and a pair of internal control primers are added to the bottom of each PCR reaction plate, and the last reaction well contains B27 universal primers To determine the negative or positive of the sample; all primers are freeze-dried at the bottom of the PCR reaction plate.

[0034] The distribution of 13 pairs of specific amplification primers in the PCR reaction plate is as follows:

[0035]

[0036]

[0037] A pair of internal control primers were also added to each well to amplify a 600bp product, and the sequences of the internal control primers were as follows:

[0038] 5'-TGCATCTGGACATGCTTGCT-3'

5'-TGGCTGGAGGAGACTCCAAA-3'

[0039] 3) PCR reagents

[0040] DNA polymer...

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Abstract

The invention provides a human HLA-B27 gene typing kit. The human HLA-B27 gene typing kit comprises a PCR amplification primer, DNA polymerase and a reaction buffer solution, wherein the DNA polymerase is a hot initiated Taq polymerase which is modified by polyethylene glycol (PEG) and can tolerate PCR reaction inhibitors in blood. The invention also provides a nucleotide sequence of the PCR amplification primer. The human HLA-B27 gene typing kit has the technical effects that the adopted DNA polymerase is the hot initiated Taq polymerase which is modified by PEG and can tolerate PCR reaction inhibitors, PCR amplification on whole blood of human (nucleic acid extraction does not need to be carried out) can be realized, PCR amplification is directly carried out by virtue of blood, and DNA extraction does not need to be carried out; the human HLA-B27 gene typing kit can realize HLA-B27 negative and positive identification on a sample; meanwhile, a gene subtype result of an HLA-B27 positive sample can be obtained, and 24 HLA-B27 gene subtypes and other gene subtypes of 13 HLAB sites can be detected by every eight holes (eight linked pipes).

Description

technical field [0001] The invention relates to a human HLA-B27 genotyping kit, which belongs to the technical field of genetic engineering. Background technique [0002] Human leukocyte antigen (Human Leukocyte Antigen, HLA) is located in the 21.31 region of the short arm of chromosome 6, which is the region with the highest gene density and the most abundant polymorphism in the known human chromosome, and is divided into HLA-Ⅰ, Ⅱ and Ⅲ Gene. HLA-B27 is a classic HLA-class I molecule, expressed on the surface of all nucleated cells and platelets, with a high degree of polymorphism, known to have 105 subtypes, named HLA-B*27:01~HLA-B* 27:105. [0003] Ankylosing spondylitis (AS) is an autoimmune disease of unknown cause that mainly occurs in young and middle-aged men, mainly chronic inflammation of axial joints, and the most common clinical manifestations are waist pain and stiffness. The total incidence of AS in my country is 0.3%, about 4.5 million cases, more men than ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2521/101C12Q2527/125
Inventor 陈炤源王浩
Owner JIANGSU WEIHE BIOTECH
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