Detection of beta-thalassemia mutations using a multiplex amplification refractory mutation system assay

Area of Science:

• Molecular Biology
• Genetics
• Medical Diagnostics

Background:

• Beta-thalassemia is a significant public health concern, particularly in Thailand.
• Accurate and efficient identification of beta-thalassemia mutations is crucial for genetic counseling and prenatal diagnosis.
• Existing diagnostic methods may not cover the spectrum of prevalent mutations effectively.

Purpose of the Study:

• To develop and evaluate a multiplex amplification refractory mutation system (M-ARMS) assay for detecting common beta-thalassemia mutations in the Thai population.
• To assess the sensitivity and specificity of the M-ARMS assay in a cohort of individuals with beta-thalassemia trait.

Main Methods:

• Development of two sets of multiplex amplification refractory mutation system (M-ARMS) assays targeting specific common beta-thalassemia mutations.
• Application of the M-ARMS assays to 282 individuals with beta-thalassemia trait.
• Confirmation of mutations using DNA sequencing for cases not detected by M-ARMS.

Main Results:

• The M-ARMS assay demonstrated high detection rates, with 97.8% of individuals testing positive for beta-thalassemia trait.
• Set 1 M-ARMS assay identified 91.5% of cases, primarily detecting codons 41/42, codon 17, IVS-I-1, and codons 71/72 mutations.
• Set 2 M-ARMS assay identified 6.4% of cases, detecting -28, -87, and IVS-II-654 mutations; 2.2% required further sequencing.

Conclusions:

• The developed M-ARMS assay is a robust, accurate, simple, speedy, and cost-effective tool for beta-thalassemia mutation analysis in Thailand.
• This assay significantly facilitates genetic counseling and prenatal diagnosis for severe thalassemia in high-risk pregnancies.
• The M-ARMS assay provides a valuable method for identifying a broad range of prevalent beta-thalassemia mutations.