The application of the amplification refractory mutation system (ARMS)‎ for characterization β-thalassemia mutations in Duhok

Abstract EN

Background and Objectives β-thalassemias are important health problems in Duhok governorate, requiring establishing proper control programs, based in screening, counseling and prenatal diagnosis.

Such programs should be reliable and cost effective.

Amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) is the cornerstone of such programs worldwide.

However, such technology has not been yet applied in our country.

The aim of the study is introducing ARMS-PCR technique and applying it to the molecular characterization of β-thalassemia in the province.

Methods A total of 68 unrelated patients with thalassemia major/intermedia were screened for eight main thal mutations namely : IVS-II-I (G>A), Codon 44 (-C), IVS-I-1 (G>A), Codon 5 (-CT), IVS-I-6 (T>C), IVS-I-5 (G>C), Codon 39 (C>T) and Codon 8/9 (+G) sequentially, using ARMS-PCR method.

Uncharacterized cases were further evaluated using Multiplex PCR-Reverse hybridization.

Results ARMS-PCR allowed the full characterization of 85.3%.

If ARMS is followed by RH then 97% of cases will be fully characterized.

The most common mutation identified were IVS-II-1, Codon 44, IVS-I-6 and Codon 39.

In 40 cases where PCR-Reverse hybridization as well as ARMS were available the concordance rate was 100%.

Conclusion The use of ARMS-PCR for the above eight mutations is a feasible, cost effective and a reliable technique and should be advocated as the first line molecular diagnostic tool for characterization of β-thal as part of our regional control program for this disorder.