Comparison of high-resolution melting analysis to denaturing high performance liquid chromatography in the detection of point mutations in MEFV, F5, and F2 genes.

BACKGROUND/AIM Sensitive and cost-effective detection of point mutations is important in genetics research. Denaturing high-performance liquid chromatography (DHPLC) is known to be one of the most sensitive techniques for point mutation detection. A more recent technique, high-resolution melting (HRM), is based on the melting behavior of PCR products. In this study, the efficiency and sensitivity of HRM and DHPLC for the detection of MEFV, F5, and F2 gene point mutations were evaluated. MATERIALS AND METHODS We studied 15 patients with MEFV mutations (E148Q, M680I, M694V, or V726A), 7 patients with the F51691G>A mutation, and 12 patients with the F220210G>A mutation. All mutations were screened by HRM and DHPLC. RESULTS All mutations were successfully detected by HRM. However, only 4 (MEFVE148Q and M680I, F51691G>A, and F220210G>A) of 6 mutations were successfully detected with DHPLC. CONCLUSION Our study showed that HRM is more sensitive than DHPLC for detection of the studied point mutations.

[1]  J. Qian,et al.  Development of high-resolution melting analysis for the detection of the MYD88 L265P mutation. , 2013, Clinical biochemistry.

[2]  Gyeong Suk Kim,et al.  DHPLC is a highly sensitive and rapid screening method to detect BRAF(V600E) mutation in papillary thyroid carcinoma. , 2013, Experimental and molecular pathology.

[3]  Ş. Şahin,et al.  Distribution of prothrombin G20210A, factor V Leiden, and MTHFR C677T mutations in the middle Black Sea area (Tokat) of Turkey , 2012, Turkish Journal of Medical Sciences.

[4]  P. Conesa‐Zamora,et al.  Comparison of allelic discrimination by dHPLC, HRM, and TaqMan in the detection of BRAF mutation V600E. , 2011, The Journal of molecular diagnostics : JMD.

[5]  Shih-Cheng Chang,et al.  High-resolution melting analysis is a more effective approach for screening TSC genes mutations. , 2011, Genetic testing and molecular biomarkers.

[6]  C. Ayuso,et al.  Comparison of high-resolution melting analysis with denaturing high-performance liquid chromatography for mutation scanning in the ABCA4 gene. , 2010, Investigative ophthalmology & visual science.

[7]  S. Julia,et al.  Validation of high-resolution DNA melting analysis for mutation scanning of the LMNA gene. , 2009, Clinical biochemistry.

[8]  G. Millat,et al.  Rapid, sensitive and inexpensive detection of SCN5A genetic variations by high resolution melting analysis. , 2009, Clinical biochemistry.

[9]  B. McCrindle,et al.  Detection of familial defective apoB (FDB) mutations in hypercholesterolemic children and adolescents by denaturing high performance liquid chromatography (DHPLC). , 2008, Clinical biochemistry.

[10]  David M. Thomas,et al.  High resolution melting analysis for the rapid and sensitive detection of mutations in clinical samples: KRAS codon 12 and 13 mutations in non-small cell lung cancer , 2006, BMC Cancer.

[11]  Salwa Khan,et al.  Hereditary thrombophilia , 2006, Thrombosis journal.

[12]  A. Bakkaloğlu,et al.  Familial Mediterranean Fever (FMF) in Turkey: Results of a Nationwide Multicenter Study , 2005, Medicine.

[13]  Carl T Wittwer,et al.  Sensitivity and specificity of single-nucleotide polymorphism scanning by high-resolution melting analysis. , 2004, Clinical chemistry.

[14]  K. Murphy,et al.  Mutation and single nucleotide polymorphism detection using temperature gradient capillary electrophoresis , 2003, Expert review of molecular diagnostics.

[15]  Carl T Wittwer,et al.  High-resolution genotyping by amplicon melting analysis using LCGreen. , 2003, Clinical chemistry.

[16]  R. Gershoni-baruch,et al.  Familial Mediterranean fever: prevalence, penetrance and genetic drift , 2001, European Journal of Human Genetics.

[17]  Isabelle Touitou,et al.  The spectrum of Familial Mediterranean Fever (FMF) mutations , 2001, European Journal of Human Genetics.

[18]  P. Oefner,et al.  Denaturing high‐performance liquid chromatography: A review , 2001, Human mutation.

[19]  N. Akar,et al.  MEFV mutations in Turkish patients suffering from familial Mediterranean fever , 2000, Human mutation.

[20]  P. Oefner,et al.  Denaturing high-performance liquid chromatography detects reliably BRCA1 and BRCA2 mutations. , 1999, Genomics.

[21]  S N Thibodeau,et al.  Denaturing high performance liquid chromatography (DHPLC) used in the detection of germline and somatic mutations. , 1998, Nucleic acids research.

[22]  Jacques Demaille,et al.  A candidate gene for familial Mediterranean fever , 1997, Nature Genetics.

[23]  P. Reitsma,et al.  A common genetic variation in the 3'-untranslated region of the prothrombin gene is associated with elevated plasma prothrombin levels and an increase in venous thrombosis. , 1996, Blood.

[24]  Pieter H. Reitsma,et al.  Mutation in blood coagulation factor V associated with resistance to activated protein C , 1994, Nature.

[25]  P. Cerutti,et al.  Genotypic mutation analysis by RFLP/PCR. , 1993, Mutation research.

[26]  L. Lerman,et al.  Comprehensive detection of single base changes in human genomic DNA using denaturing gradient gel electrophoresis and a GC clamp. , 1990, Genomics.

[27]  E. Sohar,et al.  Familial Mediterranean fever. , 1955, Klinische Padiatrie.

[28]  F. Rivier,et al.  Validation of high-resolution DNA melting analysis for mutation scanning of the CDKL5 gene: identification of novel mutations. , 2013, Gene.

[29]  R. Lai,et al.  Detection of c-KIT and PDGFRA gene mutations in gastrointestinal stromal tumors: comparison of DHPLC and DNA sequencing methods using a single population-based cohort. , 2010, American journal of clinical pathology.

[30]  Robert Palais,et al.  Simultaneous mutation scanning and genotyping by high-resolution DNA melting analysis , 2007, Nature Protocols.

[31]  R. Margraf,et al.  Mutation scanning of the RET protooncogene using high-resolution melting analysis. , 2006, Clinical chemistry.

[32]  E. Gross,et al.  Identification of specific BRCA1 and BRCA2 variants by DHPLC. , 2000, Human mutation.

[33]  A. Nataraj,et al.  Single‐strand conformation polymorphism and heteroduplex analysis for gel‐based mutation detection , 1999, Electrophoresis.

[34]  B. Dahlbäck,et al.  Thrombophilia as a multigenic disease. , 1999, Haematologica.