Severe hemophilia A disease-causing gene mutation detection kit

Abstract

The invention belongs to the field of biotechnology and biomedicine, and particularly relates to a kit for detecting inversion mutation of intron 22 and intron 1 of a severe hemophilia A disease-causing gene. The severe hemophilia A disease-causing gene mutation detection kit comprises Inv22 PCR (polymerase chain reaction) liquid and Inv1 PCR liquid, the Inv22 PCR liquid comprises three primers for detecting gene inversion of the intron 22 of the severe hemophilia A disease-causing gene, and the Inv1 PCR liquid comprises three primers for detecting gene inversion of the intron 1 of the severe hemophilia A disease-causing gene. Long fragment amplification stability of Inv22, detection sensitivity, PCR product quantity and electrophoresis detection abundance are improved, the problems of short preservation time, poor temperature tolerance and the like of reagents are solved by further adding Taq DNA (deoxyribonucleic acid) polymerase inhibitors, and stability of clinical application of the kit is improved.

Description

technical field [0001] The invention belongs to the field of biotechnology and biomedicine, in particular to a kit for detecting the inversion mutation of intron 22 and intron 1 of the severe hemophilia A pathogenic gene. Background technique [0002] Hemophilia A (hemophilia A, HA), also known as hemophilia A, is the most common X-linked recessive hereditary bleeding disorder. deficiencies or functional deficiencies. Patients with HA often suffer from spontaneous or traumatic bleeding, and bleeding can occur in any location, but joint bleeding is the most common. Repeated joint bleeding can lead to disability and even life-threatening of patients. Clinically, HA is divided into heavy, medium and light according to the procoagulant activity of plasma FVIII and the severity of symptoms, and 50% of them are severe. [0003] The incidence of HA in my country is about 1 / 5000 in the male population. Hemophilia is divided into hemophilia A and hemophilia B (hemophilia B, HB) a...

AI Technical Summary

Problems solved by technology

Advantages: Simultaneous detection of hemophilia A and hemophilia B pathogenic genes, covering full-length sequences; Disadvantages: time-consuming sequencing reaction, high cost, 70%-80% of hemophilia A FVIII gene mutations are highly heterogeneous , the mutation form is complex, and the sequencing results are difficult to analyze

However, when the PCR product fragment > 400bp, the detection rate is less than 50%. To ensure the detection rate, the use of small fragment detection greatly limits the application of this method in the diagnosis of HA.

[0016] In summary, the LD-PCR method in the prior art can only detect intron 22 inversion mutations and cannot be widely used in clinical practice because the reaction system cannot meet the requirements of stability and low cost; the sequencing method cannot combine the result analysis with clinical diagnosis. Effective combination; inverse PCR method is cumbersome to operate; SSCP is only suitable for detecting point mutations, DGGE and DHPLC are cumbersome and expensive to operate, and need to be sequenced to determine the specific location of the mutation

Images