A kind of simultaneous detection kit and method of hpa and hla antigen system

Abstract

The invention provides a simultaneous detection kit and method for HPA and HLA antigen systems. By designing a probe sequence for the target gene, the separation and enrichment of the target gene can be realized, and the detection of six platelet membrane glycoprotein genes and HLA- Synchronous capture of full-length coding sequences of A, ‑B, and ‑C genes realizes simultaneous detection of HPA system and HLA‑A, ‑B, and ‑C genes in a single system, with high throughput, accurate results, and comparable detection costs With the characteristics of decline, the proportion of HLA ambiguous results has decreased significantly, realizing accurate detection, improving detection efficiency and saving detection costs. The relevant applications in the fields of clinical transfusion medicine research and genetics will be highly valued, and will play an important role in medical research units, pharmaceutical research and reagent development units.

Description

technical field [0001] The invention belongs to the technical field of genotyping detection, in particular to a synchronous detection kit and method for HPA and HLA antigen systems. Background technique [0002] Platelet transfusion is an important treatment method for clinical treatment of patients with reduced platelet count or dysfunction. However, due to the differences of alloantigens on individual platelets, platelet transfusion will be ineffective after multiple transfusions. The immune response elicited by HLA-I antigens and platelet alloantigens (HPA) on the platelet surface is the primary immune factor leading to ineffective platelet transfusion. Ineffective platelet transfusions cause difficulty in treatment and endanger the lives of patients, increasing the medical expenses and hospitalization time of patients to a certain extent; at the same time, it will increase the number and dosage of blood transfusions, which not only increases the economic burden but also ...

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

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