DHPLC screening of cystic fibrosis gene mutations

Human Mutation
M Ravnik-GlavacM Dean

Abstract

Denaturing high performance liquid chromatography (DHPLC) using ion-pairing reverse phase chromatography (IPRPC) columns is a technique for the screening of gene mutations. In order to evaluate the potential utility of this assay method in a clinical laboratory setting, we subjected the PCR products of 73 CF patients known to bear CFTR mutations to this analytic technique. We used thermal denaturation profile parameters specified by the MELT program tool, made available by Stanford University. Using this strategy, we determined an initial analytic sensitivity of 90.4% for any of 73 known CFTR mutations. Most of the mutations not detected by DHPLC under these conditions are alpha-substitutions. This information may eventually help to improve the MELT algorithm. Increasing column denaturation temperatures for one or two degrees above those recommended by the MELT program allowed 100% detection of CFTR mutations tested. By comparing DHPLC methodology used in this study with the recently reported study based on Wavemaker 3.4.4 software (Transgenomic, Omaha, NE) [Le Marechal et al., 2001) and with previous SSCP analysis of CFTR mutations [Ravnik-Glavac et al., 1994] we emphasized differences and similarities in order to refine the D...Continue Reading

References

Oct 15, 1990·Hospital Practice·F S CollinsL C Tsui
Apr 1, 1996·Nature Genetics·X Estivill
Jan 23, 1999·Clinical Chemistry and Laboratory Medicine : CCLM·L CremonesiC Gelfi
Oct 24, 2000·Kidney International·F T van der LoopH J Smeets
Feb 17, 2001·Journal of Biochemical and Biophysical Methods·R G Cotton, P J Bray
Apr 3, 2001·Genetics in Medicine : Official Journal of the American College of Medical Genetics·W W GrodyUNKNOWN Subcommittee on Cystic Fibrosis Screening, Accreditation of Genetic Services Committee, ACMG. American College of Medical Ge
Apr 3, 2001·Genetics in Medicine : Official Journal of the American College of Medical Genetics·W W Grody, R J Desnick
Jun 1, 2001·Human Mutation·W Xiao, P J Oefner
Nov 1, 2001·Molecular Diagnosis : a Journal Devoted to the Understanding of Human Disease Through the Clinical Application of Molecular Biology·R D Press
Jan 1, 2000·Pflügers Archiv : European journal of physiology·Damjan GlavačJeffrey Wine

❮ Previous
Next ❯

Citations

Sep 24, 2005·Mammalian Genome : Official Journal of the International Mammalian Genome Society·Cymbeline T CuliatQingbo Li
Jun 28, 2011·BMC Cancer·Heather L HondowAlexander Dobrovic
May 16, 2003·Clinical Chemistry and Laboratory Medicine : CCLM·Giuseppe CastaldoFrancesco Salvatore
Jan 27, 2006·The Journal of Molecular Diagnostics : JMD·Elahe ElahiYvonne R Thorstenson
Sep 19, 2007·Pediatrics International : Official Journal of the Japan Pediatric Society·Kyoko KataokaHiroyuki Tanaka
Feb 2, 2012·Immunopharmacology and Immunotoxicology·Riccardina TesseFrancesco De Robertis
Jun 24, 2014·Clinical Biochemistry·Søren Feddersen, Mads Nybo
Apr 23, 2008·Current Protocols in Human Genetics·Bing YuPeter A Underhill
Apr 24, 2003·Clinical Chemistry·Giuseppe CastaldoFrancesco Salvatore
Dec 9, 2004·DNA and Cell Biology·V Metaxa-MariatouG Nasioulas
Jan 17, 2003·Expert Review of Molecular Diagnostics·Andrew G HallJulie A E Irving
Nov 3, 2017·Mycopathologia·Frédéric GrenouilletJean-Philippe Bouchara
Mar 3, 2004·Archives of Pathology & Laboratory Medicine·Charles M Strom
Aug 30, 2003·Archives of Pathology & Laboratory Medicine·Elaine Lyon, Christine Miller

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.