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Method of detecting common ambiguous types of HLA (human leukocyte antigen) typing locus A

An ambiguous type, detection method technology, applied in the direction of microbial determination/inspection, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problem of high-resolution confirmation and accurate typing, high cost, and inability to distinguish sequences separately. Results and other issues, to achieve the effects of accurate and precise determination, cost reduction, and simple operation

Inactive Publication Date: 2019-02-19
银丰基因科技有限公司 +1
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AI Technical Summary

Problems solved by technology

One of the main reasons for the ambiguous combination of genotyping results is that the SBT method simultaneously amplifies and analyzes the sequences of two alleles. When the same sequence occurs due to a heterozygous base pair combination, The sequence results corresponding to the two alleles cannot be identified separately, that is, the ambiguous result of the same heterozygous sequence in the detection region
Most of the ambiguous results can be further confirmed by high-resolution level PCR-SSP kits, but the cost of PCR-SSP is relatively high, and as the number of alleles continues to increase, the proportion of ambiguities is also increasing, and PCR-SSP can no longer be completely resolved The problem of high-resolution confirmation of precise typing

Method used

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  • Method of detecting common ambiguous types of HLA (human leukocyte antigen) typing locus A
  • Method of detecting common ambiguous types of HLA (human leukocyte antigen) typing locus A
  • Method of detecting common ambiguous types of HLA (human leukocyte antigen) typing locus A

Examples

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Embodiment 1

[0033] Example 1 Detection method for common ambiguous types at A site of HLA typing

[0034] Proceed as follows:

[0035] (1) The ambiguous type of the A site was obtained from the HLA typing results (HLA typing was performed by the HLA SBT uTYPE 7.0 typing software, and during the routine A site typing process, some samples were found to have ambiguous types with 0 mismatch) , select the specific primers needed to distinguish the ambiguous type; there are 9 specific primers, the nucleotide sequences are shown in Table 1, and each specific primer can confirm one or more ambiguous types at the same time, each The ambiguous types that can be distinguished by the specific primers are shown in Table 1;

[0036] (2) performing sequencing PCR reaction on the purified HLA-A gene amplification product using the specific primers selected above;

[0037] (3) Purify the sequenced product, obtain the product sequence by a sequencer, compare the obtained sequence with the ambiguous type...

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Abstract

The invention discloses a method of detecting common ambiguous types of HLA (human leukocyte antigen) typing locus A. The method includes: (1) acquiring ambiguous types of locus A through HLA typing results, and selecting specific primers necessary to distinguish the ambiguous types, wherein the specific primers are shown as SEQ ID NO. 1 to 9; (2) using the selected specific primers to perform sequencing PCR (polymerase chain reaction) on the purified HLA-A gene amplification product; (3) purifying the post-sequencing product, acquiring a product sequence through a sequencer, comparing the product sequence with the ambiguous types so as to distinguish the ambiguous types, and finally determining HLA-A type information. Compared with traditional PCR-SSP (PCR-sequence specific primer) methods to determine gene ambiguous types, the method herein can accurately and precisely judge the types, has greatly reduced cost, is simple to perform, can easily provide high throughput, and is a time-saving economical option for large-scale donor HLA typing.

Description

technical field [0001] The invention relates to a detection method for common ambiguous types of HLA typing A sites and specific primers for distinguishing common ambiguous types of HLA typing A sites, belonging to the technical field of gene detection. Background technique [0002] Human leukocyte antigen (HLA) is by far the most complex genetic system in humans, with a high degree of polymorphism. Accurate HLA typing technology and typing data have been widely used in organ and hematopoietic stem cell transplantation to find HLA-matched donors, research on HLA-disease correlations, paternity identification, individual identification, and peptide-based viruses And the research and development of cancer vaccines and other application fields. The matching degree of HLA alleles between the donor and the recipient is closely related to the effect of transplantation, and accurate HLA typing can help improve the survival rate after transplantation. [0003] At present, DNA-base...

Claims

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Application Information

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IPC IPC(8): C12Q1/6881C12N15/11
CPCC12Q1/6881
Inventor 周云丽张倩鹿士振张守强
Owner 银丰基因科技有限公司
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