Gene detection kit for Hong Kong alpha-thalassemia

Abstract

The invention relates to the technical field of biology and medicament, and particularly relates to a gene detection kit for Hong Kong alpha-thalassemia. According to the invention, the gene sequence of HK alpha alpha fusion gene is further confirmed and compared with alpha-globin sequence for analysis, and primer design is carried out in a conserved sequence area of the fusion gene. The technical scheme of the invention is to provide a gene detection kit for Hong Kong alpha-thalassemia, comprising a PCR (polymerase chain reaction) solution, wherein the PCR solution contains primer HK alpha alpha-F and HK alpha alpha-R, the primer is designed aiming at the conserved sequence area of the HK alpha alpha fusion gene, and the nucleotide sequence of the conserved sequence of the HK alpha alpha fusion gene is shown in SEQ ID NO:1. The kit disclosed by the invention can be directly used for kits for HK alpha alpha detection and can specifically, quickly and stably diagnose HK alpha alpha gene type.

Description

technical field [0001] The invention relates to the technical field of biology and medicine, in particular to a kit for rapidly detecting Hong Kong-type α-thalassemia (HKαα) in clinical samples. Background technique [0002] α-Thalassemia (hereinafter referred to as α-thalassemia) is a group of genetic hemolytic hemoglobinopathies characterized by reduced or non-synthesized α-globin chains and an imbalance in the ratio of α-chain / non-α-chain. Thalassemia is one of the most common human monogenic blood diseases in the world. It is listed by the World Health Organization as six common diseases that endanger human health. It is also the most common and most harmful genetic disease in southern China. In the high-incidence areas of α-thalassemia in southern my country, the epidemiological survey of 900,000 people showed that the total incidence rate was 2.46%, and the incidence rates in Guangxi, Guangdong, Jiangxi, Sichuan and Zhejiang provinces (regions) were 14.95% respectively...

AI Technical Summary

Problems solved by technology

[0010] 1. Southern blot hybridization-restriction zymography analysis: used to be the main method for α-thalassemia gene diagnosis, but due to the cumbersome operation and long time required, it is generally only suitable for research use and not suitable for clinical application

[0011] 2. Probe Ligation Amplification Technology (MLPA): MLPA was developed in 2002. This method can detect all missing genotypes and unknown genotypes. It is highly efficient and specific, and can detect multiple in the same reaction tube. Up to 45 different nucleic acid sequence copy number changes; commonly used in the detection of α-deletion genes, but due to the high cost of detection, cumbersome operation, and long time-consuming, the detection process lasts up to 16 hours, and requires special equipment, generally only used for For research use, it is not convenient for the promotion and application of molecular screening methods

[0012] 3. Sequencing technology: The direct analysis of DNA sequences by sequencing technology has always been considered as the gold standard for clinical testing; at present, sequencing technology is facing the problem of standardization and certification of sample processing (nucleic acid extraction) amplification, signal detection, and specific errors in sequencing platforms and error correction problems, as well as possible problems in bioinformatics, clinical verification procedures and standardization problems, etc., have not been effectively resolved, so they have not been widely used in clinical practice.

In the detection of thalassemia, the reports are limited to the detection of one genotype, and the workload is heavy. Since the probe needs to be fluorescently labeled, the detection cost is high

[0016] 7. PCR-oligonucleotide probe (ASO) method: This technology can quickly and easily detect known mutations of the thalassemia gene (α T and beta T ), with high sensitivity and accuracy, but one hybridization can only detect one mutation, and also requires isotope probes, which is difficult to popularize and apply

[0021] 3. In the existing patented technology of α-thalassemia detection products, there is no detection directly for HKαα

[0022] 4. At present, the detection of HKαα thalassemia is based on clinical phenotype, with the help of existing thalassemia detection kits combined with genetic analysis of parental genotypes. There is no fast and direct method for detecting HKαα thalassemia

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