Efficient 21-hydroxylase deficiency related gene detection method

Abstract

The invention discloses an efficient 21-hydroxylase deficiency related gene detection method. The method comprises the following steps: S1, extracting genome DNA of a sample to be detected; s2, aiming at point mutation on the CYP21A2 gene to be detected, determining a sequence of a CYP21A2 gene segment according to GenBank in NCBI (National Center of Biotechnology Information), designing three pairs of specific primers through software, and carrying out three PCR (Polymerase Chain Reaction) amplification; s3, performing long-fragment PCR amplification on the genome DNA to obtain a long-fragment amplification product; and S4, carrying out enzyme digestion on the long-fragment amplification product by using restriction enzyme, detecting the enzyme digestion product by using agarose gel electrophoresis, and carrying out high-throughput sequencing detection on the long-fragment amplification product. The detection method is efficient, sensitive, simple to operate, short in time consumption, low in cost, high in detection flux, strong in specificity, accurate and reliable in detection result and suitable for clinical popularization and application.

Description

technical field [0001] The invention relates to the technical field of molecular biology, and more specifically, relates to an efficient detection method for genes related to 21-hydroxylase deficiency. Background technique [0002] Congenital adrenal hyperplasia (CAH) is a group of diseases caused by adrenal corticosteroid synthesis disorders caused by gene mutations in essential enzymes in the process of corticosteroid synthesis. It is the most common inherited metabolic disease, 21-hydroxylase deficiency 21-hydroxylase deficiency (21-OHD) is the first discovered, most studied and most common congenital adrenal hyperplasia, accounting for 90-95% of CAH, is an autosomal recessive genetic disease of cortisol synthesis disorder . Human 21-hydroxylase (CytochromeP450c21, CYP21), also known as CYP21 or P450c21, is one of the key enzymes for the body to synthesize cortisol. It is a cytochrome P450 enzyme that is only expressed in adrenal tissue. (or) Mineralocorticosteroids dec...

AI Technical Summary

Problems solved by technology

At present, the diagnosis of 21-OHD mainly relies on clinical symptoms and biochemical tests. Due to the lack of specificity and sensitivity of this diagnosis, 21-OHD is prone to misdiagnosis and missed diagnosis, and this method cannot analyze the genotype of patients.

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