Affiliations: Department of Biotechnology, University of Turku,
FIN-20520 Turku, Finland | Department of Virology, University of Turku, FIN-20520
Turku, Finland
Note: [] Correspondence: Minna Sjöroos, Åbo Akademi
University, BioCity, 3rd floor, Tykistökatu 6 A, FIN-20520 Turku, Finland
Tel: +358 2 2154018; Fax: +358 2 2154745 E-mail: minna.sjoroos@utu.fi
Abstract: A microtitration plate based time-resolved fluorescence (TRF)
hybridisation assay was developed for HLA typing utilising biotinylated
sequence-specific catching probes and europium (Eu) labelled gene
locus-specific detection probe to allow time-resolved fluorometer reading of
the reaction. In an application for HLA-DQA typing a 228 base pair long region
of the polymorphic exon 2 of DQA1 gene was amplified and the denatured PCR
product distributed into streptavidin-coated microtitration wells together with
the detection probe and one of the catching probes. After incubation and
washes, the enhancement solution was added and specific hybridisation signal
detected by measuring the emitted light. A series of 100 isolated genomic DNA
samples were studied using biotinylated probes specific for DQA1*01,
*0101/0104, *0103/0201/0601, *0201, *03, *0401/0601, *05 and *0502 alleles with
results demonstrating the capacity of the test to detect aimed alleles. A
series of whole blood spot samples were also studied and the results confirmed
the applicability of this modification of the test.
