title: An automated PCR platform with homogeneous time-resolved fluorescence detection and dry chemistry assay kits.
abstract: We have developed a novel instrument platform, GenomEra, for small-scale analysis of nucleic acids. The platform combines a rapid thermal cycler, an integrated time-resolved fluorescence measurement unit, and user-friendly software for the analysis of results. Disposable low-cost plastic reaction vessels are designed specifically for the instrument and contain all of the assay-specific reagents in dry form. The appropriate assay protocol is specified on barcodes printed under the vessels and is automatically initiated by the software. Detection is based on the use of sequence-specific probes labeled with intrinsically fluorescent europium or terbium chelates and complementary quencher probes, which enable sensitive, homogeneous closed-tube assays without the risk of carryover contamination. The detection limit of the instrument (background + 3 SD) is approximately 20 pmol/L for both chelates with a dynamic range of nearly four orders of magnitude. The functionality of the platform is demonstrated with a dual-label homogeneous polymerase chain reaction (PCR) assay for the detection of Salmonella using a Magda CA Salmonella assay kit. An internal amplification control is included in each reaction to eliminate false negative results caused by PCR inhibition. Qualitative assay results are automatically interpreted by the software and are available 45 min after sample addition.
authors.affiliation: Abacus Diagnostica, FI-20520 Turku, Finland. virve.hagren@abacusdiagnostica.com
authors.forename: Virve
authors.initials: V
authors.lastname: Hagren
authors.forename: Piia
authors.initials: P
authors.lastname: von Lode
authors.forename: Anniina
authors.initials: A
authors.lastname: Syrjälä
authors.forename: Tero
authors.initials: T
authors.lastname: Soukka
authors.forename: Timo
authors.initials: T
authors.lastname: Lövgren
authors.forename: Hannu
authors.initials: H
authors.lastname: Kojola
authors.forename: Jussi
authors.initials: J
authors.lastname: Nurmi
mesh_terms: D001331:Automation; D004209:Disposable Equipment; D005453:Fluorescence; D007753:Laboratories; D008163:Luminescent Measurements / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D012680:Sensitivity and Specificity; D012984:Software; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013997:Time Factors