A type I neurofibromatosis pathogenic mutation gene and an etiological diagnostic reagent based on the mutation gene

Abstract

The invention relates to neurofibromatosis type I virulence gene mutation and an etiological diagnosis agent based on neurofibromatosis type I virulence gene mutation. The etiological diagnosis agent comprises point mutation analysis kits including a total RNA extraction system kit, a RNA reverse transcription and cDNA amplification system kit and a cDNA sequencing system kit. The total RNA extraction system kit is used for total RNA extraction of peripheral venous blood of a tested subject; the cDNA sequencing system kit is used for PCR amplification of cDNA obtained by reverse transcription and five primer pairs as shown in SEQ ID NO.7-16, nested PCR amplification and sequencing of cDNA five long segments obtained by amplification and primer pairs as shown in SEQ ID NO.17-60, and comparison with an original NF1 gene coding DNA sequence; if c.7106G>A and p.W2369X mutation exists in an amplified product, a neurofibromatosis type I patient is determined. The etiological diagnosis agent has advantages of time saving, accuracy and low test cost.

Description

technical field [0001] The invention relates to the fields of molecular biology and gene detection, in particular to a type I neurofibromatosis pathogenic mutation gene and an etiology diagnostic reagent based on the mutation gene. Background technique [0002] Neurofibromatosis type I (Neurofibromatosis type I, NF1) was first reported by von Recklinghausen in 1882, so it is also called von Recklinghausen disease. Type I neurofibromatosis is generally autosomal dominant, some sporadic. Clinically, the main manifestations are skin coffee spots and multiple soft fibrous tumors of the skin; freckles and iris melanoma hamartomas in non-exposed parts (such as armpits and groin); multiple organs and systems can also be involved, such as eyes, bones, blood vessels , endocrine system, central and peripheral nervous system, etc., and may be accompanied by mental retardation, congenital dysplasia, and extremely rare malignant transformation. [0003] The disease is caused by mutatio...

AI Technical Summary

Problems solved by technology

[0006] Since the NF1 gene contains 58 constitutive exons and 3 alternatively spliced ​​exons, the traditional sequencing analysis method for each exon takes a long time, and it usually takes six or seven days to complete the detection of each sequencing sample

Other detection methods, such as NGS, FISH or MLPA, etc., need to be equipped with expensive large-scale instruments and equipment, and require high experimental conditions, resulting in a significant increase in detection costs

[0007] In addition, in addition to point mutations, there are many types of mutations in the NF1 gene, such as large deletions, duplication / deletion of one or more exons, etc.

It is easy to miss the detection of NF1 gene mutation by a single method, which is also a difficult problem for clinical genetic diagnosis personnel in the detection of NF1 pathogenic gene mutation

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