title: Modeling real-time PCR kinetics: Richards reparametrized equation for quantitative estimation of European hake (Merluccius merluccius).
abstract: Real-time PCR is the most sensitive method for detection and precise quantification of specific DNA sequences, but it is not usually applied as a quantitative method in seafood. In general, benchmark techniques, mainly cycle threshold (Ct), are the routine method for quantitative estimations, but they are not the most precise approaches for a standard assay. In the present work, amplification data from European hake (Merluccius merluccius) DNA samples were accurately modeled by three sigmoid reparametrized equations, where the lag phase parameter (λc) from the Richards equation with four parameters was demonstrated to be the perfect substitute for Ct for PCR quantification. The concentrations of primers and probes were subsequently optimized by means of that selected kinetic parameter. Finally, the linear correlation among DNA concentration and λc was also confirmed.
authors.affiliation: Grupo de Bioquı́mica de Alimentos, Instituto de Investigacións Mariñas (CSIC), Vigo, Spain. asanchez@iim.csic.es
authors.forename: Ana
authors.initials: A
authors.lastname: Sánchez
authors.forename: José A
authors.initials: JA
authors.lastname: Vázquez
authors.forename: Javier
authors.initials: J
authors.lastname: Quinteiro
authors.forename: Carmen G
authors.initials: CG
authors.lastname: Sotelo
mesh_terms: D000465:Algorithms; D000818:Animals; D030801:Animals, Genetically Modified / Q000378:metabolism*; D004247:DNA / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism; D005513:Food Inspection / Q000379:methods*; D030841:Food, Genetically Modified; D048388:Gadiformes / Q000378:metabolism*; D007700:Kinetics; D008962:Models, Theoretical; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017747:Seafood / Q000032:analysis*
chemical_list: D029941:Fish Proteins; D004247:DNA