title: Identification of G6PD Mediterranean mutation by amplification refractory mutation system.
abstract: 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.
authors.affiliation: Laboratorio di Biochimica Clinica, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy. maffi@iss.it
authors.forename: Donatella
authors.initials: D
authors.lastname: Maffi
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Pasquino
authors.forename: Patrizia
authors.initials: P
authors.lastname: Caprari
authors.forename: Maria Pia
authors.initials: MP
authors.lastname: Caforio
authors.forename: Paolo
authors.initials: P
authors.lastname: Cianciulli
authors.forename: Francesco
authors.initials: F
authors.lastname: Sorrentino
authors.forename: Maria Pia
authors.initials: MP
authors.lastname: Cappabianca
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Salvati
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D005260:Female; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics*; D006801:Humans; D008297:Male; D019083:Mediterranean Region; D009154:Mutation / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D005954:Glucosephosphate Dehydrogenase