P1732 Evaluation of an Automated Sample Preparation System (mdx) in Combination with Affigene CMV Trender
International journal of antimicrobial agents(2007)
Abstract
Genomic DNA extraction for genotyping analysis is performed from blood samples and is time consuming. We describe a more rapid DNA extraction method, “DBS-miniMAG”, that combines filter paper dried blood spots (DBS) with the NucliSens® miniMAG™ semi-automated instrument (bioMérieux). To assess the performance of this method, a post-PCR HLA-DR shared epitope (SE) oligotyping assay was used as a read-out in a cohort of 72 arthritis patients. This new method was compared to the standard manual DBS extraction protocol using FTA® reagents (Whatmann Bio-Science), and to a reference phenol-chloroform-based method using EDTA whole blood samples. Higher yield of PCR amplicons was observed with DNA extracts obtained using “DBS-miniMAG” method. The intra- and inter-assay variability of the “DBS-miniMAG” method was similar to that obtained with “DBS-FTA” washing process. Concerning the HLA-DR SE genotyping, “DBS-miniMAG” and “DBS-FTA” methods gave 100% concordance compared to the reference phenol-chloroform method. More importantly, the hands-on time and the turnaround time for “DBS-miniMAG” were both two-times shorter than for “DBS-FTA” protocol. Therefore, the “DBS-miniMAG” combination could facilitate polymorphism analysis in routine clinical practice and the creation of large DNA banks using very small amounts of blood.
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Diagnosis
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