Methylene blue as an electrochemical indicator for DF508 cystic fibrosis mutation detection
2010 (English)In: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 396, no 4, 1423-1432 p.Article in journal (Refereed) Published
Cystic fibrosis is one of the most common lifeshortening,childhood-onset inherited diseases. Among the1,000 known cystic fibrosis-related mutations, DF508 is themost common, with a frequency varying between 50% and70% according to geographical areas and populationtypology. In this work, we report the use of methyleneblue as an electrochemical reporting agent in the discriminationof synthetic PCR analogue of the DF508 cysticfibrosis mutation (Mut) from the wild type (Wt). Atoptimum experimental condition, a discrimination factorbetween mutant and wild type of approximately 1.5-foldwas found. The proposed assay was quantitative and linearin the range of 10–100 nM, exhibiting a limit of detectionof 2.64 nM. Electrochemical studies at variable ionicstrength conditions allowed further elucidation of themechanism of the methylene blue (MB)–DNA interaction.To the best of our knowledge, this is the first report ofdetection of hybridisation solely via guanine-specific MB–DNA interaction simultaneously in MB solution, independentof electrostatic interaction as demonstrated in the ionicstrength study. The introduction of formamide in thehybridization buffer, to improve discrimination, was alsoinvestigated. Finally, mutant wild type discrimination wasdemonstrated, at 10 nM concentration, with the use of amulti-sensor setup.
Place, publisher, year, edition, pages
Springer Publishing Company, 2010. Vol. 396, no 4, 1423-1432 p.
Methylene blue . Cystic fibrosis . DF508 . DNA sensor . Electrochemistry
IdentifiersURN: urn:nbn:se:liu:diva-92115DOI: 10.1007/s00216-009-3369-5OAI: oai:DiVA.org:liu-92115DiVA: diva2:620163