Kit for detecting abnormal hemoglobin gene mutation

Abstract

The invention discloses a kit for detecting human abnormal hemoglobin gene mutation. The kit for detecting abnormal hemoglobin gene mutation comprises a reagent A for detecting eight abnormal alpha-globin gene point mutations and a reagent B for detecting six abnormal beta-globin gene point mutations, the reagent A comprises a group of alpha globin specific primer pairs and eight alpha globin mutation site specific probes; the reagent B comprises two groups of beta globin specific primer pairs and five beta globin mutation site specific probes. The kit for detecting abnormal hemoglobin gene mutation can simultaneously detect 14 kinds of abnormal hemoglobin gene mutation, and is more comprehensive in detection site, higher in efficiency, high in accuracy and good in repeatability.

Description

technical field [0001] The application relates to the technical field of human abnormal hemoglobin gene detection, in particular to a kit for detecting abnormal hemoglobin gene mutations. Background technique [0002] The pathogenesis of abnormal hemoglobin is a group of hereditary hemoglobinopathy in which the molecular structure of hemoglobin changes due to changes in the primary structure of globin. The internal amino acids of normal hemoglobin are non-polar amino acids, which constitute the sites where subunits contact each other, heme and globin chains contact, and globin chain helical segments contact; if amino acid abnormalities occur inside the molecule, the hemoglobin space The impact of conformation and function is greater, and the clinical manifestations are obvious. Some abnormal hemoglobin can cause thalassemia phenotype, and its common pathological mechanism is that the expression of affected globin chains is reduced by changes in peptide chain structure or ge...

AI Technical Summary

Problems solved by technology

Among them, the amplification hindered mutation system PCR (ARMS-PCR) uses two PCR reaction tubes to detect each mutation site one by one. False Negative and False Positive Results

Allele-specific oligonucleotide hybridization (ASO) can reduce false negative and false positive results, but still needs to detect each mutation site one by one, the detection throughput is low, time-consuming and laborious

PCR-Reverse Dot Blot (PCR-RDB) method, this method has the advantages of high sensitivity, good specificity and high throughput, and can detect three mutation types of WS, QS and CS at the same time. It is widely used in the detection of α-thalassaemia gene, but its manual operation is cumbersome. The α2 globin gene is first amplified by PCR, and then the detection results are observed through a series of operations such as denaturation, hybridization, membrane washing, and color development. The detection time is relatively short. long, reducing work efficiency

At the same time, the opening operation of PCR products also greatly increases the risk of contamination in the laboratory

DNA sequencing technology is to amplify by PCR first, then purify the PCR products, construct a sequencing library, perform sequencing on the machine, and perform data analysis on the sequencing results, etc.; there are also problems such as cumbersome operations, long time-consuming, and contamination

[0008] In general, the existing methods for the detection of abnormal hemoglobin gene point mutations have high detection costs, heavy workload, cumbersome operations, and low detection throughput. It is difficult to achieve automation and standardization, and cannot meet the current large-scale population screening and clinical The need for routine testing; and, currently there is no kit on the market that can more comprehensively detect abnormal hemoglobin

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