title: Molecular assay optimized by Taguchi experimental design method for venous thromboembolism investigation.
abstract: Two mutations - Factor V Leiden (1691G > A) and the 20210G > A on the Prothrombin gene - are key risk factors for a frequent and potentially fatal disorder called Venous Thromboembolism. These molecular alterations can be investigated using real-time Polymerase Chain Reaction (PCR) with Fluorescence Resonance Energy Transfer (FRET) probes and distinct DNA pools for both factors. The objective of this paper is to present an application of Taguchi Experimental Design Method to determine the best parameters adjustment of a Molecular Assays Process in order to obtain the best diagnostic result for Venous Thromboembolism investigation. The complete process contains six three-level factors which usually demands 729 experiments to obtain the final result, if using a Full Factorial Array. In this research, a Taguchi L27 Orthogonal Array is chosen to optimize the analysis and reduce the number of experiments to 27 without degrading the final result accuracy. The application of this method can lessen the time and cost necessary to achieve the best operation condition for a required performance. The results is proven in practice and confirmed that the Taguchi method can really offer a good approach for clinical assay efficiency and effectiveness improvement even though the clinical diagnostics can be based on the use of qualitative techniques.
authors.affiliation: Sao Paulo State University, UNESP, Department of Production Engineering, Guaratinguetá, SP, Brazil. hcelani@uol.com.br
authors.forename: Helder Jose
authors.initials: HJ
authors.lastname: Celani de Souza
authors.forename: Cinthia B
authors.initials: CB
authors.lastname: Moyses
authors.forename: Fabrício J
authors.initials: FJ
authors.lastname: Pontes
authors.forename: Roberto N
authors.initials: RN
authors.lastname: Duarte
authors.forename: Carlos Eduardo
authors.initials: CE
authors.lastname: Sanches da Silva
authors.forename: Fernando Lopes
authors.initials: FL
authors.lastname: Alberto
authors.forename: Ubirajara R
authors.initials: UR
authors.lastname: Ferreira
authors.forename: Messias Borges
authors.initials: MB
authors.lastname: Silva
mesh_terms: D000465:Algorithms; D004252:DNA Mutational Analysis / Q000379:methods*; D005163:Factor Analysis, Statistical; D005165:Factor V / Q000235:genetics*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005838:Genotype; D006801:Humans; D009154:Mutation; D011516:Prothrombin / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D054556:Venous Thromboembolism / Q000175:diagnosis / Q000235:genetics
chemical_list: D005165:Factor V; D011516:Prothrombin