Rapid single-step FCGR3A genotyping based on SYBR Green I fluorescence in real-time multiplex allele-specific PCR.

Abstract:

:A polymorphism at position 559 in the cDNA of the FCGR3A gene encoding the FcgammaRIIIa generates two allotypes with either a valine (Val) or a phenylalanine (Phe) at amino acid position 158. This polymorphism is of major importance in immunopathology and in pharmacogenetics, especially monoclonal antibody treatments. In this study, we report a single-step and single-tube method for FCGR3A-158V/F genotyping by real-time multiplex allele-specific PCR and melting curve analysis in the presence of SYBR Green I fluorescent dye. Results obtained from 113 samples showed 100% accuracy compared to those obtained with a conventional PCR-based allele-specific restriction assay. Although this method requires expensive equipment, it is inexpensive in terms of consumables. It is also very rapid, reliable and suitable for large-scale analysis.

journal_name

J Immunol Methods

authors

Dall'Ozzo S,Andres C,Bardos P,Watier H,Thibault G

doi

10.1016/s0022-1759(03)00123-6

keywords:

subject

Has Abstract

pub_date

2003-06-01 00:00:00

pages

185-92

issue

1-2

eissn

0022-1759

issn

1872-7905

pii

S0022175903001236

journal_volume

277

pub_type

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