Blotting membrane strip, PCR primer and kit for diagnosis of PAH gene mutation

Abstract

The invention discloses a blotting membrane strip, a PCR primer and a kit for diagnosis of PAH gene mutation. The blotting membrane strip comprises a substrate, and a mutation detection probe which is fixed on the substrate. Each mutation detection probe is separately corresponding to one PAH gene mutation site, and the PAH gene mutation detection sites comprise the following sites: R111X, R176X, EX6-96A>G, R241C, R243Q, R252Q, Y356X, V399V and R413P; mutation detection probe comprises a sequence including 15-25 bases, and the middle position of the sequence comprises a mutant base corresponding to the PAH gene mutation site; the mutation detection probe is DNA of a sequence shown in SEQ ID No.1-SEQ ID No.9 or a reverse complement sequence. The blotting membrane strip, the PCR primer and the kit have the advantages of high flux, high efficiency, low price, usage convenience and the like, and are good for realizing clinic rapid detection and large scale population screening.

Description

technical field [0001] The invention relates to molecular biology detection, in particular to a hybrid membrane strip for diagnosing phenylalanine hydroxylase (phenylalanine hydroxylase, PAH) gene mutation and a polymerase chain reaction for amplifying gene fragments of samples to be tested (Polymerase Chain Reaction, abbreviated as PCR) primers and a kit for diagnosing PAH gene mutation. Background technique [0002] Phenylketonuria (PKU) is an autosomal recessive disorder of phenylalanine metabolism that seriously threatens human health, most of which (98%-99%) are caused by phenylalanine hydroxylase (phenylalaninehydroxylase, PAH) gene defect, resulting in the decrease or lack of PAH activity, resulting in the abnormal metabolism of phenylalanine, the accumulation of alanine and its bypass metabolites in blood and tissues, and finally leading to irreversible damage to the central nervous system. Damage, and symptoms such as abnormal mental behavior, epilepsy and mental r...

AI Technical Summary

Problems solved by technology

[0005] At present, traditional gene diagnosis methods include enzyme digestion, restriction fragment length polymorphism (RFLP), direct sequencing, etc. These methods are either not qualitative, or time-consuming and labor-intensive, and require expensive equipment and consumables. Unfortunately, these methods are difficult to detect multiple gene mutation sites at the same time

Although the gene chip method can detect different gene loci at the same time, it requires expensive equipment and consumables, and is not suitable for large-scale clinical promotion, and its application is limited.

Images