Cormoran® Direct HLA-B27 Genotyping PCR Kit

HLA-B27 detection directly from blood

The Cormoran® Direct HLA-B27 Genotyping PCR Kit enables qualitative detection of HLA-B27 directly from diluted EDTA whole blood, without a separate DNA extraction step. Purified genomic DNA can alternatively be used as the starting material.

The kit combines a hot-start Genotyping PCR Master Mix with a target-specific Primers & Probes Mix for hydrolysis probe-based real-time PCR.

Two target regions with an internal control

The assay targets HLA-B27-specific sequences in exon 2 and exon 3 of the HLA-B gene. Both target regions contribute to the HLA-B27 signal in the FAM channel.

An internal amplification control is detected simultaneously in the ROX channel. This control helps identify reactions affected by PCR inhibition or insufficient amplifiable DNA, supporting interpretation of negative HLA-B27 results.

The assay provides a qualitative HLA-B27 result and does not differentiate individual HLA-B27 subtypes.

Key advantages

• Direct use of diluted EDTA whole blood without DNA extraction.

• Compatible with purified genomic DNA.

• HLA-B27 detection and internal control in the same reaction.

• Hot-start formulation for convenient reaction setup at room temperature.

• Real-time fluorescence detection without gel electrophoresis or melting-curve analysis.

• Two-component reagent system for 20 µL reactions.

Simple workflow

Prepare the sample according to the manual. Combine 4 µL Genotyping PCR Master Mix, 6 µL HLA-B27 Primers & Probes Mix and 10 µL prepared sample.

Run the specified real-time PCR program and evaluate amplification in the FAM and ROX channels. A real-time PCR instrument equipped with both detection channels is required.

Based on published research

The HLA-B27 assay design is based on the primer and probe sequences described by Geiger et al. (2020), with Axel Muendlein as senior author, for real-time PCR-based HLA-B27 screening directly in whole blood.

Reference: Geiger K, Zach C, Leiherer A, Fraunberger P, Drexel H, Muendlein A. Real-time PCR based HLA-B*27 screening directly in whole blood. HLA. 2020;95(3):189–195.  https://doi.org/10.1111/tan.13767

Storage and shipping

Shipped without cooling. Store at −20 °C immediately upon receipt. Protect the HLA-B27 Primers & Probes Mix from light.

For research use only. Not for use in diagnostic procedures.