A homogeneous HLA-B*27 genotyping assay using dried reagent mixtures

Article type: Research Article

Authors: Kiviniemi, Minna | Ilonen, Jorma^; | Lövgren, Timo

Affiliations: Immunogenetics Laboratory, University of Turku, Tykistökatu 6 A, 4th floor, FIN-20520 Turku, Finland | Department of Clinical Microbiology, University of Kuopio and Biocenter Kuopio, FIN-70211, Kuopio, Finland | Department of Biotechnology, University of Turku, Tykistökatu 6 A, 6th floor, FIN-20520 Turku, Finland

Note: [] Corresponding author: M. Kiviniemi. Tel.: +358 2 333 7009; Fax: +358 2 3337000; E-mail: minna.kiviniemi@utu.fi

Abstract: The presence of HLA-B*27 allele with patients suspected with ankylosing spondylitis can be used in the diagnostic process. We have developed an assay for typing for the HLA-B*27 in whole blood dried on sample collection cards using pre-dried reagent wells and homogeneous time-resolved fluorescence based PCR approach. Essentially only the sample needs to be added to the dry ready-to-use reaction well in order to start the homogenous amplification assay. The method was validated with 229 samples also typed with an existing DELFIA-based method and results of both assays were 100% concordant. The dried reagents were shown to be stable at least up to eight weeks at room temperature without any decline in their performance.

Keywords: Dry chemistry, HLA-B*27, homogeneous assay, PCR

DOI: 10.3233/DMA-2009-0653

Journal: Disease Markers, vol. 27, no. 2, pp. 85-91, 2009