1. Introduction
Phenylketonuria (PKU) is an autosomal recessive metabolic disease caused
by mutations in the gene encoding phenylalanine hydroxylase (PAH) (1).
At present, the early diagnosis and intervention of PKU through neonatal
disease screening can avoid the damage to the nervous system and other
tissues caused by PAH mutations (2). At the same time, prenatal
diagnosis can reduce the chance of giving birth to children with PKU in
high-risk families. From the perspective of the three-level prevention
system, the most effective way to prevent PKU is to identify carriers
before pregnancy, so as to take effective measures to avoid the birth of
PKU children (3). However, due to the variety and heterogeneity of PAH
gene mutations, a lot of basic and clinical research is needed to avoid
the birth of children with PKU.
As the incidence of PKU and the spectrum of PAH gene mutations in
Shaanxi province, China are not clear, it is necessary to combine PKU
prevention strategies with the screening and diagnosis of PAH gene
mutations, so as to form an effective three-level prevention model and
improve the efficiency of PKU prevention. For gene diagnosis of PKU, the
most commonly used detection technologies at present mainly include DNA
sequencing, multiplex ligation-dependent probe amplification (MLPA),
short tandem repeat linkage analysis, etc. However, there are problems
associated with the use of these approaches, such as complex operation,
high cost, high requirements for laboratory personnel, and not being
conducive to popularization (4-5). Therefore, the purpose of this study
was to develop amplification refractory mutation system-polymerase chain
reaction (ARMS-PCR) combined with fluorescent probe technology to detect
the mutation hotspots of the PAH gene, in order to develop a PAH gene
screening kit that is convenient, fast, cheap, accurate, and suitable
for all levels of screening agencies, thereby providing technical
support for the “one-level prevention” of PKU (3).