Identification of G6PD Mediterranean mutation by amplification refractory mutation system

• Published in: Clinica chimica acta 2002-07, Vol.321 (1), p.43-47
• Main Authors: Maffi, Donatella, Pasquino, Maria Teresa, Caprari, Patrizia, Caforio, Maria Pia, Cianciulli, Paolo, Sorrentino, Francesco, Cappabianca, Maria Pia, Salvati, Anna Maria
• Format: Article
• Language: English
• Subjects: Amplification refractory mutation system; Biological and medical sciences; DNA Mutational Analysis - methods; Female; G6PD Mediterranean mutation; Glucose-6-phosphate dehydrogenase deficiency; Glucosephosphate Dehydrogenase - genetics; Hematology; Humans; Investigative techniques, diagnostic techniques (general aspects); Male; Medical sciences; Mediterranean Region; Mutation - genetics; Nucleic Acid Amplification Techniques - methods; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; PCR amplification; Polymerase Chain Reaction; Sensitivity and Specificity
• ISSN: 0009-8981; 1873-3492
• DOI: 10.1016/S0009-8981(02)00098-0

Background: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common X chromosome-linked hereditary enzymopathy in humans. The authors have developed an amplification refractory mutation system (ARMS) to detect the G6PD Mediterranean mutation (nt. 563 C→T) that is the most frequent among Caucasian population. Methods: Specific forward polymerase chain reaction (PCR) primers, within exon 6 of the G6PD gene, were designed: ARMS M complementary to the mutated DNA sequence and ARMS N complementary to the wild-type DNA. They were paired with a common reverse primer. The new method was validated using known DNA samples from 72 G6PD-deficient patients carrying the G6PD Mediterranean mutation ascertained by the restriction enzyme analysis. The ARMS test was performed on DNA extracted both from blood or saliva samples. Results: The ARMS test showed an excellent reproducibility and a complete concordance with the endonuclease cleavage reference method. At the same time, it is more rapid and less expensive. Conclusions: The described molecular test may be a method of choice to identify the G6PD Mediterranean mutation. It could also be helpful to obtain a definite diagnosis of G6PD Mediterranean heterozygotes, which is not feasible by using red blood cell enzyme activity measurements.