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Rapid amplification diagnostic kit and amplification method for HLA (human leucocyte antigen) high-resolution genetic locus

A diagnostic kit and gene locus technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of increasing the economic burden of the testee, requiring a large amount of testing samples, and long testing time, etc. Achieve the effect of small sample amount, good accuracy and high specificity

Inactive Publication Date: 2018-09-25
新开源博畅(武汉)生物科技有限公司
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

Existing gene diagnosis mainly adopts traditional sequencing method or gene chip detection method, which requires a large amount of test samples, and the detection time is long and the steps are cumbersome, which greatly increases the economic burden of the testee

Method used

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  • Rapid amplification diagnostic kit and amplification method for HLA (human leucocyte antigen) high-resolution genetic locus
  • Rapid amplification diagnostic kit and amplification method for HLA (human leucocyte antigen) high-resolution genetic locus
  • Rapid amplification diagnostic kit and amplification method for HLA (human leucocyte antigen) high-resolution genetic locus

Examples

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Embodiment Construction

[0029] The rapid amplification diagnostic kit and amplification method of HLA high-resolution gene loci provided by the present invention will be further described in detail and completely in conjunction with the examples below. The embodiments described below are exemplary only for explaining the present invention and should not be construed as limiting the present invention.

[0030] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples were purchased from the market unless otherwise specified.

[0031] The instruments and reagents adopted in the present invention are as follows:

[0032] PTC-255 PCR amplification instrument, MJ Research, USA;

[0033] Pyrosequencer, Wuhan First Biotechnology Co., Ltd.;

[0034] TaqDNA polymerase, dNTPs (10mmol / L), 10×Buffer and MgCl 2(25mmol / L) (Dalian TaKaRa company); AmpliTaqGold polymerase (AppliedBiosystems company); KlenowFra...

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Abstract

The invention discloses a rapid amplification diagnostic kit and amplification method for HLA (human leucocyte antigen) high-resolution genetic locus. The kit comprises DNA polymerase and an amplification primer; the HLA high-resolution genetic locus is HLA-DRB1, having a template sequence shown as in sequence table SEQ ID NO: 1; the amplification primer is used for performing linear index PCR (polymerase chain reaction) amplification; an upstream sequence of the amplification primer is shown as in sequence table SEQ ID NO: 2, and a downstream sequence of the amplification primer is shown as in sequence table SEQ ID NO: 3. A target fragment amplification method employing linear index amplification has the advantages that the amplification primer that is designed precisely has high synergistic efficiency and low mismatching rate, amplification efficiency is greatly improved, single-stranded DNA can be amplified directly to serve for pyrosequencing, performing marking and dual-strand separation on an amplification product is not required, complex experimental steps are omitted, and damage of strong-basicity reagents to amplified fragments is lessened.

Description

technical field [0001] The invention relates to a rapid amplification diagnostic kit and amplification method for HLA high-resolution gene loci, belonging to the biological field, in particular to the biological detection field. Background technique [0002] Human leukocyte antigen (HLA) is the human major histocompatibility antigen complex (MHC), the main component of specific immune recognition and immune response in the body, and one of the key factors affecting the success of hematopoietic stem cell transplantation. Reliable HLA gene Frequency, especially high-resolution gene frequency data, is an important basis for predicting the possibility of finding an HLA-matched donor, and it also has important application value for HLA-disease correlation research, population genetics research, and forensic science research. At present, there have been many reports on HLA in different regions and different races in the Chinese population, but most of them are based on low-resolut...

Claims

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Application Information

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IPC IPC(8): C12Q1/6881C12Q1/6806C12Q1/6869
CPCC12Q1/6806C12Q1/6869C12Q1/6881C12Q2600/156C12Q2531/113C12Q2565/301
Inventor 郑超陈素华吴祚达
Owner 新开源博畅(武汉)生物科技有限公司
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