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Identification of G6PD Mediterranean mutation by amplification refractory mutation system
Donatella Maffi1, Maria Teresa Pasquino, Patrizia Caprari
1Laboratorio di Biochimica Clinica, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy. maffi@iss.it
Summary
A new molecular test accurately detects the common G6PD Mediterranean mutation. This amplification refractory mutation system (ARMS) offers a faster, cheaper, and more reliable method for diagnosing Glucose-6-phosphate dehydrogenase deficiency.
Area of Science:
- Genetics
- Molecular Biology
- Biochemistry
Background:
- Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a prevalent X-linked hereditary enzymopathy.
- The G6PD Mediterranean mutation (nt. 563 C-->T) is the most frequent variant in Caucasian populations.
Purpose of the Study:
- To develop and validate an amplification refractory mutation system (ARMS) for detecting the G6PD Mediterranean mutation.
- To provide a molecular diagnostic tool for G6PD deficiency.
Main Methods:
- Designed specific polymerase chain reaction (PCR) primers for mutated (ARMS M) and wild-type (ARMS N) G6PD sequences.
- Validated the ARMS test using DNA from 72 G6PD-deficient patients with the Mediterranean mutation.
- Applied the ARMS test to DNA extracted from both blood and saliva samples.
Main Results:
- The ARMS test demonstrated excellent reproducibility.
- Achieved complete concordance with the established endonuclease cleavage reference method.
- The ARMS method proved to be more rapid and cost-effective than the reference method.
Conclusions:
- The developed ARMS test is a suitable method for identifying the G6PD Mediterranean mutation.
- This molecular test can aid in the definitive diagnosis of G6PD Mediterranean heterozygotes, which is challenging with enzyme activity assays.