Synchronous detection kit and method for HPA and HLA antigen systems

Abstract

The invention provides a synchronous detection kit and method for HPA and HLA antigen systems. By designing a probe sequence aiming at a target gene, separation and enrichment of the target gene are achieved, synchronous capture of six platelet membrane glycoprotein genes and full-length coding sequences of HLA-A, -B and -C genes is achieved, and the HPA system and the HLA-A, -B and -C genes are synchronously detected in a single system. The synchronous detection kit and method have the advantages of high throughput, accurate result and reduced detection cost on year-on-year basis, the proportion of ambiguous results for HLA is obviously reduced, accurate detection is realized, the detection efficiency is improved, and the detection cost is saved. The related application of the synchronous detection kit and method in the fields of clinical blood transfusion medical research, genetics and the like is highly paid attention to, and the synchronous detection kit and method play an important role in medical research units, pharmaceutical research units and reagent development units.

Description

technical field [0001] The invention belongs to the technical field of genotyping detection, and in particular relates to a simultaneous detection kit and method for HPA and HLA antigen systems. Background technique [0002] Platelet transfusion is an important treatment for patients with decreased platelet count or dysfunction in clinical practice. However, due to the differences in alloantigens on individual platelets, platelet transfusion may be ineffective after multiple transfusions. The immune response triggered by HLA class I antigen on platelet surface and platelet alloantigen (HPA) is the primary immune factor leading to ineffective platelet transfusion. Ineffective platelet transfusion causes difficult treatment and endangers the life of the patient, and increases the patient's medical expenses and hospitalization time to a certain extent; at the same time, it will increase the number and amount of blood transfusion of the patient, which not only increases the econ...

AI Technical Summary

Problems solved by technology

The PCR-SSP method is a low-throughput detection technology. The detection of each HPA antigen system needs to be amplified and measured in a separate reaction well, which is a heavy workload and is not suitable for routine detection; the PCR-SSO method based on the Luminex platform, its probe Most of the design refers to the Caucasian data, and does not cover the unique antigen system of the Chinese population; PCR-SBT method can achieve accurate sequencing, but the detection method is mainly based on Sanger sequencing technology, each HPA system needs to be designed and amplified separately, and the operation is cumbersome The problem of low quality and insufficient throughput limits the detection of large-scale samples; at the same time, these HPA typing methods are only used to detect known polymorphisms, the range of base sequences determined is limited, and it is difficult to achieve high-throughput detection; HLA-A, -B, -C gene detection mainly uses PCR-SBT and Luminex PCR-SSO method, PCR-SBT mainly analyzes the polymorphism of the 2nd, 3rd, 4th exons, and each gene requires multiple reactions Or multiple experiments can be completed, and there are some ambiguous HLA allele combinations in the sequencing, PCR-SBT technology cannot accurately determine these allele combinations, which is not conducive to the rapid and accurate identification of HLA genotypes

The Luminex PCR-SSO method is based on known polymorphic sites for detection, which can only achieve low and medium resolution capabilities, and the typing results also have a high frequency of ambiguous gene combinations, which cannot achieve accurate typing

In addition, the existing HPA and HLA genotyping techniques are to detect a single gene separately. Since each gene contains multiple exons, it needs multiple PCR amplification and sequencing reactions to complete. The method of typing HPA antigen system and HLA-A, -B, -C genes is heavy and time-consuming, which greatly affects the efficiency of detection, and it is difficult to meet the needs of rapid and accurate identification of HPA and HLA genotypes

Images