Author/Authors :
Dastsooz, Hassan shiraz university of medical sciences - Department of Medical Genetics, Department of Molecular Medicine, ايران , Imanieh, Mohammad Hadi shiraz university of medical sciences - Shiraz Transplant Research Center, Gastroenterohepatology Research Center, Namazi Teaching Hospital, ايران , Dehghani, Mohsen shiraz university of medical sciences - Shiraz Transplant Research Center, Gastroenterohepatology Research Center, Namazi Teaching Hospital, ايران , Haghighat, Mahmood shiraz university of medical sciences - Shiraz Transplant Research Center, Gastroenterohepatology Research Center, Namazi Teaching Hospital, ايران , Moini, Maryam shiraz university of medical sciences - Gastroenterology and Hepatology Research Center - Department of Internal Medicine, ايران , Fardaei, Majid shiraz university of medical sciences - Stem Cell and Transgenic Technology Research Center - Department of Medical Genetics, Department of Molecular Medicine, ايران
Abstract :
Background: Wilson disease is a rare disorder of copper metabolism due to mutation in ATP7B gene. Proper counseling of patients with Wilson disease, and their families necessitates finding mutation in ATP7B gene. Finding mutations in ATP7B gene with 21 exons, and more than 500 mutations is expensive and time-consuming. Objectives: The aim of this study was to provide a simple multiplex amplification refractory mutation system PCR (M-ARMS-PCR) for screening eight common mutations in ATP7B gene. Patients and Methods: Two sets of ARMS mutant and normal specific primer pairs were designed for genotyping of p.R778L, p.R969Q, p.H1069Q, and p.3400delC mutations as Set 1 and p.W779G, c.3061-1G A, p.I1102T, and p.N1270S mutations as Set 2. The Multiplex ARMS assay was then subsequently tested in 65 patients with Wilson disease with known and unknown ATP7B mutations. Results: Using these two sets, we identified H1069Q mutation in four patients, c.2335T G mutation in three, c.3061-1G A splice site mutation in five, c.3305T C mutation in one, and c.3809A G mutation in two patients. Conclusions: The Multiplex ARMS assay used in this study can be an efficient, reliable, and cost effective method as a primary screen for patients with Wilson disease.
