کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1966771 1538732 2008 4 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Rapid diagnosis of Wilson disease by a 28-mutation panel: Real-time amplification refractory mutation system in diagnosing acute Wilsonian liver failure
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Rapid diagnosis of Wilson disease by a 28-mutation panel: Real-time amplification refractory mutation system in diagnosing acute Wilsonian liver failure
چکیده انگلیسی

BackgroundWilson disease is one of the commonest inherited and potentially fatal yet treatable liver disorders. About 5–27% patients present with acute liver failure and require prompt chelation therapy and life-saving liver transplantation. Diagnosis during acute liver failure is particularly difficult with short time allowance. Direct molecular diagnosis remains the most decisive tool but is often hindered by demanding techniques and numerous mutations. We developed a one-step, 3-h, reproducible, and accurate real-time amplification refractory mutation system which can simultaneously detect 28 ATP7B mutations.MethodsPrimers were designed to complement the mutant sequence at the 3′ end. The mutations were p.S105X, p.Q511X, p.R616Q, p.S693P, p.S693C, p.R778L, p.A874V, p.T888P, p.R919G, p.T935M, p.P992L, p.M1025R, p.D1047V, p.I1148T, p.R1156H, p.T1178A, p.V1216M, p.P1273Q, p.G1281C, p.R1320S, p.V1334D, p.V176SfsX28, p.G869EfsX4, IVS3+1G>T, IVS4-1G>C, IVS4-5T>G, IVS6+9A>G, and IVS9+5G>T. Reaction was performed using QuantiTect SYBR Green PCR Master Mix on an Applied Biosystems StepOne thermal cycler. Values of the threshold cycle were compared between normal and mutant alleles.ResultsPrimers of all mutations were highly specific with absence of wild-type amplification. All the results were validated by direct DNA sequencing.ConclusionsThis rapid and cost-efficient method allows wide mutation coverage, rendering the SYBR-green assay feasible and attractive for large-scale routine application.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Clinica Chimica Acta - Volume 398, Issues 1–2, December 2008, Pages 39–42
نویسندگان
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