Comparison of Amplification Refractory Mutation System and Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Techniques Used for the Investigation of MEFV Gene Exon 10 Point Mutations in Familial Mediterranean Fever Patients Living in Çukurova Region (Turkey)

Abstract

Familial Mediterranean fever (FMF) is an autosomal recessive inherited disease characterized by recurrent fever, serositis and arthritis. The disease is highly prevalent in Mediterranean basin populations. Recently, the gene responsible for FMF (MEFV) was cloned and at least 40 MEFV gene mutations have been identified. The most frequently observed mutations in the MEFV gene are M694V, M694I, M680I, and V726A. These occur within exon 10 of the gene, and account for 85% of the known MEFV alleles. In this study, the reliability and economical aspects of amplification refractory mutation system (ARMS) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) techniques were compared for analyzing the frequencies of the major point mutations of 90 unrelated patients with FMF from the Cukurova region in Turkey. Both techniques yielded similar results: The ratio of independent alleles of 90 patients carrying one of the tested mutations was 81.1%; patients consisted of 12 different genotypes. In 64 of 90 patients (71.1%) mutations were observed in both alleles. Thirty-six patients (40%) were homozygous for the same mutation, 28 (31.1%) were heterozygous for different mutations. Eighteen patients (20%) were heterozygous for one allele with one of the four mutations but the other allele was unknown. In 8 patients (8.8%) no mutation could be detected. The most frequently observed mutation was M694V (51.66%), followed by M680I (17.22%), V726A (10.55%), and M694I (1.66%). In conclusion ARMS and PCR-RFLP techniques were equally reliable to detect the mutations in Turkish FMF patients. However, the ARMS technique was found to be more rapid and economical than the PCR-RFLP techniques.

Frequency of MEFV gene mutations in Hatay province, Mediterranean region of Turkey and report of a novel missense mutation (I247V)

“…M694I is a rare mutation which has a frequency between 0% and 2.8% in the MEFV alleles in Turkish, and Iranian Azerbaijani Turks patients with FMF (Akar et al, 1999;Akin et al, 2010;Bonyadi et al, 2009;Ceylan et al, 2012;Demirkaya et al, 2008;Dogan et al, 2012;Dundar et al, 2011;Gulec et al, 2012;Gunesacar et al, 2005;Ulgenalp, 2009;Yesilada et al, 2012). In our study, the frequency of this mutation was consistent with previous reports in the literature.…”

MEFV Gene Mutations in a Sample of Turkish Population: A Retrospective Study

“…After MEFV gene was cloned, several methods like PCR-RFLP, ARMS, and DGGE have been used in mutation detection in FMF patients. 15,16 In a study which is held in Turkey, PCR-RFLP and RH results of the patients are compared and RH is found as a recent method used in FMF mutation screening. 16 Briefly, the method involves multiplex PCR amplification of target regions on DNA.…”

“…15,16 In a study which is held in Turkey, PCR-RFLP and RH results of the patients are compared and RH is found as a recent method used in FMF mutation screening. 16 Briefly, the method involves multiplex PCR amplification of target regions on DNA. Biotin-labeled PCR products are hybridized to oligonucleotide probes on a nylon membrane colored enzymatically afterwards.…”

“…Biotin-labeled PCR products are hybridized to oligonucleotide probes on a nylon membrane colored enzymatically afterwards. 15,16 As an advantage of RH, in addition to the frequent mutations, seldom mutations can be screened at the same time. In different populations, the frequencies of these mutations are reported between 0-5%.…”

Screening for Familial Mediterranean Fever M694V and V726A Mutations in the Greek Population

“…DNA samples were screened for the two most common MEFV mutations, M694V and V726A, using the Amplification Refractory Mutation System (ARMS) (Eisenberg et al, 1998;Gunesacar et al, 2005). The sequences of the polymerase chain reaction (PCR) primers used for the detection of mutations M694V and V726A are shown in Table 1.…”