QTYPE is a Real Time PCR assay that utilizes the multiplexing capabilities of hydrolysis probes to increase the capacity for unique assays in each well that enables the determination in a low resolution HLA-A, B, C, DRB1, DRB3, DRB4, DRB5, DQA1, DQB1, DPA1 and DPB1 alleles. Using hydrolysis probes allows quick and robust typing from low input DNA concentrations and removes the risk of misinterpreting primer dimers and other PCR artefacts seen in Real Time techniques based on intercalating dyes.
Due to their sequence-specific nature, hydrolysis probes also contribute towards increased specificity of the assay, potentially decreasing the number of ambiguities in the reported results.
Each position of the QTYPE 384 well plate contains prealiquoted dried primers as well as a dual-labelled hydrolysis probe designed to bind to a location between the primers. Each primer pair binds to a specific subset of alleles. Also a Master Mix including Taq Polimerase enzime is included with the kit.
QTYPE products use up to four separate detection channels per well to determine whether a reaction is positive or negative. One of these is reserved for an internal amplification control, designed to amplify a conserved region of the human genome, and used to ensure that DNA was added to each well. Currently, two channels are used for HLA-specific reactions.
QTYPE enables speed and precision in HLA typing at a low to intermediate resolution for samples that require a fast turn-around-time and has been validated on Real Time LightCycler 480 II (Roche) and Quantstudio™ 6/7 y ViiA™-7 (Thermo Fisher) devices.
