Method for detecting PAH (Polycyclic Aromatic Hydrocarbon) gene mutation by utilizing high-resolution melting curve analysis technique

Abstract

The invention discloses a method for detecting PAH (Polycyclic Aromatic Hydrocarbon) gene mutation by utilizing a high-resolution melting curve analysis technique. The method is characterized by comprising the following steps: adopting a silica gel adsorption method to extract the genome DNA of an oral epithelial cell / peripheral blood cell sample of a detected person, designing the third, sixth, seventh, eleventh and twelfth exon sequences of the PAH gene as detection primers of a template, and carrying out mutation detection on the gene sequences after PCR (Polymerase Chain Reaction) amplification according to the high-resolution melting curve analysis technique. The method has the advantages of high sensitivity, good specificity and high detection speed and can quickly screen common mutation sites of phenylalanine hydroxylase genes. The invention provides a new basis for the method for detecting PAH gene mutation.

Description

technical field [0001] The invention relates to a detection method for phenylalanine hydroxylase gene mutation, which belongs to the technical field of molecular biology. Background technique [0002] The phenylalanine hydroxylase (PAH) gene is located on chromosome 12q23.2, with a total length of about 90kb, including 13 exons and 12 introns. The length of the exon is a single strand of 1353bp, the mRNA is translated into an enzyme monomer containing 451 amino acids, and the monomer is polymerized into a functional PAH enzyme. PAH gene mutations can lead to liver phenylalanine hydroxylase deficiency, and phenylalanine cannot be converted to tyrosine normally, resulting in metabolic disorders in the liver, leading to phenylketonuria (PKU) in children. Phenylketonuria is an autosomal recessive genetic disease with abnormal amino acid metabolism. The incidence rate in Europe and the United States is 1 / 10000, in my country it is 1 / 16000, and the heterozygote frequency is 1 / 50...

AI Technical Summary

Problems solved by technology

[0004] At present, the commonly used mutation detection method is direct DNA sequencing method. PCR products can be directly analyzed by DNA sequence, which can clarify the mutation site, but it has the disadvantages of time-consuming, laborious, expensive, and not suitable for detection of a large number of samples.

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