Decision criteria for rational selection of homogeneous genotyping platforms for pharmacogenomics testing in clinical diagnostics.
BACKGROUND
Genotyping is crucial for the identification of genetic markers underlying the development of neoplastic diseases and for determining individual variations in response to specific drugs. Technologies which can accurately identify genetic polymorphisms will dramatically affect routine diagnostic processes and future therapeutic developments in personalized medicine. However, such methods need to fulfill the principles of analytical validation to determine their suitability to assess nucleotide polymorphisms in target genes.


METHODS
This article reviews recent developments in homogeneous technologies for the genotyping of single nucleotide polymorphisms. Here, homogeneous methods essentially refer to "single-tube" assays performed in a liquid phase. For the appropriate choice of any method, several criteria must be considered: 1) detection of known genetic variations; 2) analytical performance including specificity, sensitivity and robustness of the method; 3) availability of large platforms and required equipment; 4) suitability of platforms and tests for routine diagnostics; 5) suitability for high throughput implementation.


BACKGROUND
This review is intended to provide the reader with an understanding of these various technologies for pharmacogenomic testing in the routine clinical laboratory. A brief overview is provided on the available technologies for the detection of known mutations, a specific description of the homogeneous platforms currently employed in genotyping analysis, and considerations regarding the proper assessment of the analytical performance of these methods. Based on the criteria proposed here, potential users may evaluate advantages and limitations of the various analytical platforms and identify the most appropriate platform according to their specific setting and diagnostic needs.
Di Francia
Raffaele
R
Hematology-Oncology and Stem Cell Transplantation Unit, National Cancer Institute, Fondazione G. Pascale IRCCS, Naples, Italy. rdifrancia@iapharmagen.org
Frigeri
Ferdinando
F
Berretta
Massimiliano
M
Cecchin
Erika
E
Orlando
Claudio
C
Pinto
Antonio
A
Pinzani
Pamela
P
D031541:Fluorescence Resonance Energy Transfer; D005838:Genotype; D006801:Humans; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D020135:Peptide Nucleic Acids / Q000737:chemistry / Q000378:metabolism; D010597:Pharmacogenetics; D020641:Polymorphism, Single Nucleotide; D057285:Precision Medicine; D011786:Quality Control; D017422:Sequence Analysis, DNA
D009841:Oligonucleotides; D020135:Peptide Nucleic Acids; C477371:locked nucleic acid