Chip, amplification reagent and kit for directly and simultaneously detecting alpha-thalassemia and beta-thalassemia mutation sites

Abstract

The invention relates to a chip, an amplification reagent and a kit for directly and simultaneously detecting alpha-thalassemia and beta-thalassemia mutation sites, and belongs to a molecular diagnosis technology. According to the present invention, based on direct multiplex PCR, Gap-PCR and reverse dot blot combined detection principle, the corresponding amplification primers and the corresponding probes are designed according to the mutation or deletion sites of each genotype, the primer is labeled with biotin, the probe is labeled with amino, a gene chip is used as substrate, the probe is immobilized on the DNA chip, the PCR product amplified by the specific primer is hybridized with the probe, and the diagnosis of thalassemia is performed by interpreting a signal coloring box.

Description

technical field [0001] The invention relates to a molecular diagnosis technique for α and β thalassemia, in particular to a kit for simultaneously detecting 6 kinds of α thalassemia mutations and 19 kinds of point mutation β thalassemias in combination with direct PCR (DirectPCR) hybridization technology. Background technique [0002] Thalassemia (Thalassemia, referred to as thalassemia) is one of the most common genetic diseases in the world, which is caused by the reduction or absence of synthesis of one or more peptide chains of the globin chain composed of hemoglobin (Hb) tetramer. According to the adult major globin chain (α 2 beta 2 ), the disease is divided into α-thalassemia and β-thalassemia. Affected individuals mainly originate from the Mediterranean region, the Middle East, Transcaucasus, Central Asia, the Indian subcontinent, Southeast Asia, and East Asia. The incidence is high in places south of the Yangtze River in my country such as Guangdong, Guangxi, Hain...

AI Technical Summary

Problems solved by technology

[0008] In addition, these techniques require DNA extraction, which is cumbersome and complicated, time-consuming, prone to cross-contamination, and experiments require DNA automatic extraction instruments, precision instruments and reagent consumables, which are expensive and cannot be widely used clinically.

The quality of DNA is a very critical factor for the success of PCR, and the process of extracting and purifying DNA is easily contaminated, making the experiment prone to false negatives or false positives, which can easily cause missed or false detections

[0009] In addition, these techniques require a large number of samples, and cold chain equipment is necessary for the transportation of samples collected in remote areas. A three-level packaging system is required, and the storage space for samples is large; the biosecurity risk factor is high

There is no patented technology for direct detection of thalassemia in whole blood, amniotic fluid and DBS, and there is no such product on the market

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