Human SLC25A13 gene detection kit and application

Abstract

The invention discloses a human SLC25A13 gene detection kit and an application thereof. The human SLC25A13 gene detection kit comprises c.851854deL4, c.16381660dup, IVS6 + 5Ggt, c.851854deL4, c.16381660dp, IVS6 + 5Ggt, a, IVS 16ins3kb, IVS 4ins6kb, IVS 11 + 1Ggt, IVS 4ins6kb, IVS 11 + 1Ggt, IVS 3gt, a, c, 1399Cgt, c, 1399Cgt; t, and c.1078Cgt; the primer group and the probe group are related to eight related loci such as T, T and the like; wherein a competitive anti-slip design is adopted in the design process of the probe at the c.16381660dup site, so that a nucleic acid combination 2 at the site can effectively distinguish heterozygote and homozygous mutation at the c.16381660dup site; the kit provided by the invention is reliable in typing detection result, the consistency rate of the kit and direct sequencing reaches 100%, the sensitivity of the kit is higher than that of a traditional sequencing method, and the kit is simple and rapid to operate and beneficial to large-scale popularization.

Description

technical field [0001] The invention relates to the technical field of genotyping detection, in particular to a human SLC25A13 gene detection kit and its application. Background technique [0002] Citrin deficiency disease (Citrin Deficiency, CD) is an autosomal recessive genetic disease caused by SLC25A13 biallelic mutation, and neonatal intrahepatic cholestasis caused by Citrin deficiency (NeonataLIntrahepaticChoLestasis caused by Citrin Deficiency, NICCD ) is currently the most important pediatric CD phenotype. [0003] The incidence of neonatal intrahepatic cholestasis is about 1 / 2500-1 / 5000, which is one of the most common types of liver diseases in children and ranks second among high-risk patients with inherited metabolic diseases in China. According to reports, the carrier rate of the SLC24A13 gene mutation in the Chinese population is as high as 1 / 63, while that in the southern population is as high as 1 / 48. With the Yangtze River as the boundary, the mutation car...

AI Technical Summary

Problems solved by technology

[0005] The symptoms of NICCD are temporary and complex, and it is difficult to establish clear clinical diagnostic indicators. At present, there are no recognized clinical and biochemical diagnostic criteria for NICCD at home and abroad. SLC25A13 gene mutation analysis is considered to be a reliable means of diagnosing NICCD. Both SLC25A13 alleles have pathogenic mutations to confirm the diagnosis of the disease

[0006] At present, domestic and foreign studies have shown that traditional analysis methods such as PCR-RFLP, LA-PCR DNA, and Sanger sequencing have high requirements for samples, complex technical equipment, long detection time and high cost, and are difficult to carry out on a large scale.

Chinese patent document CN103421909A aims at the above problems, and adopts fluorescent PCR to detect mutations of SLC25A13 gene-related sites. The innovation of this patent is that it is easy to operate and low in cost. The disadvantage is that the result interpretation is prone to false negative and false positive results.

Chinese patent document CN108048553A adopts a more traditional Sanger sequencing method to detect mutations in SLC25A13 gene-related sites. The innovation of this type of method is that there are many detection sites, and the disadvantage is high cost.

Chinese patent document CN108753952A has been upgraded on the basis of CN108048553A, adopts the method of multiplex PCR-sequencing, can detect a plurality of SLC25A13 gene-related loci in one PCR and sequencing process, has reduced cost, but still has complicated operation, consumption time issue

The next-generation sequencing developed on the basis of Sanger sequencing has also been applied to the detection of mutations related to the SLC25A13 gene, such as the Chinese patent document CN108913761A. Low degree of localization of instruments

Chinese patent document CN111073959A has successfully screened several gene mutations involved in Citrin deficiency by using fluorescent PCR melting curve method. Requirements for sample quality, and there is a risk of melting curve drift. At the same time, the patent has lower clinical sensitivity and specificity due to fewer detection sites.

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