Kit for detecting Rett syndrome pathogenic gene SNP loci based on next generation sequencing and detection method thereof

Abstract

The invention discloses a kit for detecting Rett syndrome pathogenic gene SNP loci based on next generation sequencing and a detection method thereof and belongs to the field of gene detection. The detection method comprises the following steps: (1) extracting sample DNA; (2) carrying out multiplex PCR reaction on a DNA template; (3) carrying out DNA fragmentation; (4) repairing DNA terminal; (5) carrying out joint connection; (6) purifying a connection product; (7) carrying out PCR enrichment; (8) purifying a PCR product; (9) detecting quality of library; (10) carrying out high-throughput sequencing; and (11) analyzing bioinformatics of sequencing results. The kit is high in detection throughput, strong in sensitivity and specificity, simple and safe to operate; the operation is more fast, accurate and complete; the kit is free of toxic and harmful substances and is free of harm to test workers and the environment.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to the detection of Rett syndrome gene mutation sites. Background technique [0002] Rett syndrome (Rett syndrome, RTT) is an X-linked neurodevelopmental disorder genetic disease, which seriously affects children's psychomotor development, and the incidence rate in girls is 1 / 10000-1 / 15000; Rett syndrome is second only to Trisomy 21 is one of the main causes of sporadic severe mental retardation; the typical clinical features of Rett are: the growth and development of infants are basically normal 6 to 18 months after birth, followed by neurodevelopmental stagnation or regression, loss of Acquired skills (such as hand function, language, etc.), slow growth in head circumference, abnormal breathing, stereotyped hand movements (such as rubbing hands, wringing hands, eating hands, etc.), accompanied by autism-like behaviors; in addition, convulsions Seizures are also one of the common sym...

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Problems solved by technology

[0004] At present, the diagnosis of Rett syndrome is mostly judged by clinical manifestations. However, it is inevitable that the diagnosis based on clinical manifestations is not convincing. Therefore, it is necessary to establish genetic testing to assist clinical early diagnosis of Rett patients; at this stage, the genetic detection of Rett Most of them are about the detection of MECP2 gene mutations, which are mainly carried out by common PCR amplification and next-generation sequencing methods. There are few samples and sites that can be detected at one time, low sensitivity, long time-consuming, and cumbersome operations; therefore, we use high-throughput sequencing to detect Suspected Rett patients are tested for MECP2, CDKL5, and FOXG1 gene mutations. Multiple mutation sites can be detected at one time, with high sensitivity, easy operation, and short time. It can quickly carry out early auxiliary diagnosis of Rett syndrome

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