Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Polymerase chain reaction-sequence based typing (PCR-SBT) method and reagent for human neutrophil alloantigen (HNA) 1-5 system gene typing

A PCR-SBT and genotyping technology, applied in the field of reagents used by the method, can solve the problems of unsystematic typing and imperfect typing methods, achieve accurate results, prevent adverse blood transfusion reactions, and prevent the occurrence of TRALI. Effect

Active Publication Date: 2014-03-05
浙江省血液中心
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current HNA antigen gene PCR-SBT typing method is not perfect enough. Although some laboratories use the PCR-SBT method to genotype individual HNA antigens, so far no systematic typing of the five HNA systems has been performed.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Polymerase chain reaction-sequence based typing (PCR-SBT) method and reagent for human neutrophil alloantigen (HNA) 1-5 system gene typing
  • Polymerase chain reaction-sequence based typing (PCR-SBT) method and reagent for human neutrophil alloantigen (HNA) 1-5 system gene typing
  • Polymerase chain reaction-sequence based typing (PCR-SBT) method and reagent for human neutrophil alloantigen (HNA) 1-5 system gene typing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] This implementation specifically takes blood donors to perform HNA1-5 system antigen genotyping as an example to describe the content of the present invention in detail. A PCR-SBT method for HNA1-5 system antigen genotyping used in the present invention specifically includes the following steps:

[0048] 1. Prepare human genomic DNA as a template for PCR amplification in subsequent steps.

[0049] Take 200 μl of whole blood to be tested, extract genomic DNA according to the instructions of the QuickGene DNA whole blood kit S kit, and use a spectrophotometer to measure the concentration and purity of the genome.

[0050] 2. Synthesize 5 pairs of amplification primers and 2 sequencing primers. For the specific sequence, please refer to the sequence in the content of the invention mentioned above, and will not repeat them. Dilute the amplification primers to 50 μM with pure water;

[0051] Prepare La-Taq enzyme (Lot: CK4501, TaKaRa), 10× buffer (Lot: CK4501, TaKaRa), Mg 2...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a polymerase chain reaction-sequence based typing (PCR-SBT) method for human neutrophil alloantigen (HNA) 1-5 system gene typing. The PCR-SBT method comprises the following steps of preparing a personal genome DNA, providing amplification primers, respectively amplifying gene sequences of HNA-1, HNA-2, HNA-3, HNA-4 and HNA-5 systems by PCR reactions, carrying out double digestion purification of the amplified products, providing sequencing primers, carrying out sequencing PCR of the purified products, carrying out sodium acetate-ethanol precipitation purification of the sequenced products, carrying out capillary electrophoresis sequencing, and carrying out software-based analysis of the sequence obtained by the previous step to determine a gene type. The invention also provides a reagent for HNA 1-5 system gene typing. Through respective sequencing of HNA 1-5 systems, the PCR-SBT method acquires an oligonucleotide sequence of a HNA gene type thereby realizing accurate typing of a HNA gene. The PCR-SBT method and the reagent have important practical meanings to medical research organizations, pharmaceutical research organizations and reagent development organizations.

Description

technical field [0001] The present invention relates to a genotyping detection method, in particular to a molecular biology detection method for human granulocyte antigen (Human Neutrophil Alloantigens, abbreviated as HNA) 1-5 system antigen genotyping, and the present invention also relates to the method Applied reagents. Background technique [0002] There are a series of blood group antigens on the human leukocyte membrane, including granulocyte-specific antigen and human leukocyte antigen. These antigens show a high degree of polymorphism in different populations. After infusion, due to differences in individual antigens, different organisms can be stimulated to produce Immune reaction can trigger certain blood transfusion adverse reactions, lead to ineffective transfusion or cause some serious diseases, so the study of granulocyte-specific antigen has important clinical significance. At present, the granulocyte antigen working group of the International Society of Bloo...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 朱发明何俊俊何吉吕杭军
Owner 浙江省血液中心
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com