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Direct HLA-B27 Genotyping PCR Kit

Product information "Direct HLA-B27 Genotyping PCR Kit"

Schnelle, Real-time Genotypisierung ohne DNA-Isolation

The HLA-B27 Direct Duplex qPCR Detection Kit enables fast and reliable qualitative detection of the HLA-B27 allele using hydrolysis probe-based real-time PCR in a duplex format. The assay is optimized for direct use of EDTA whole blood and can alternatively be performed with purified genomic DNA.

Simultaneous amplification of an HLA-B27 specific target and an internal control ensures high result reliability and minimizes the risk of false-negative outcomes. The kit is compatible with real-time PCR cyclers featuring FAM and ROX detection channels.

The Genaxxon Direct HLA-B27 Genotyping PCR Kit enables real-time genotyping without the need for prior DNA isolation, allowing for a faster and more cost-effective workflow.

Applications

  • HLA-B27 screening in suspected ankylosing spondylitis
  • Molecular support in the diagnosis of spondyloarthritides
  • Testing applications in clinical laboratories
  • Immunogenetics research applications

Key features

  • NO DNA-extraction necassary – Direct use of EDTA whole blood possible.
  • Genotyping in real-time – Rapid and efficient workflow without time-consuming DNA isolation.
  • Hot-start formulation – Enables convenient room-temperature reaction setup.
  • >Platform-independent – Compatible with all modern real-time PCR cyclers..
  • Hydrolysis probe detection (TaqMan principle). Duplex real-time PCR (target + internal control)..
  • Reliable fluorescence-based detection – Compatible with qPCR cyclers featuring FAM and ROX channels..
  • High specificity based on allele-specific primer/probe design. Two independent HLA-B27 targets (Exon 2 and Exon 3) for enhanced robustness. .
  • Eco-friendly shipping – Stable at room temperature, reducing the need for cold-chain transport.

Assay principle

The kit is based on allele-specific real-time PCR using hydrolysis probe technology. Detection is performed in a duplex format, allowing simultaneous monitoring of the target signal and an internal control. HLA-B27 detection is based on two independent HLA-B27-specific target regions within the HLA-B gene (Exon 2 and Exon 3). By targeting both regions in parallel, assay robustness is increased and the risk of false-negative results caused by rare sequence variants is reduced. The HLA-B27 signal is detected in the FAM channel. The internal control is detected in the ROX channel and ensures reliable identification of PCR inhibition or insufficient DNA release. Direct PCR from whole blood (no DNA extraction required) The kit is optimized for direct use of EDTA whole blood, enabling a simplified workflow without mandatory DNA extraction. A simple dilution of the blood sample is required. Optional is a short heat pre-treatment step (e.g., 95 °C for 10 minutes) to lyse leukocytes and release DNA. Alternatively, purified genomic DNA can be used as PCR input material.

Workflow (overview)


1. Dilute EDTA whole blood
2. Heat pre-treatment (recommended)
3. Set up the qPCR reaction
4. Run real-time PCR
5. Qualitative analysis based on amplification curves

Result interpretation

Positive: Amplification in the FAM channel (HLA-B27) with a valid internal control signal in the ROX channel.
Negative: No FAM signal, but clear amplification of the internal control in the ROX channel.
Invalid: No internal control signal (indicating PCR inhibition or incorrect sample preparation).

Cycler compatibility

A real-time PCR instrument with at least two detection channels is required: FAM and ROX.
The assay is compatible with common qPCR platforms supporting these channels.

Note

This kit is intended for screening applications. In rare cases, sequence variations of specific HLA-B27 subtypes may affect amplification performance. In case of discrepant results or specific populations, confirmatory subtyping may be recommended.

Kit contents

  • 2× HLA-B27 Direct Duplex qPCR Mastermix
  • Primer/probe mix (HLA-B27 exon targets + internal control)
  • Manual
  • Applikation / Application:

    Direct Genotyping without sample preparation

    Klassifizierungen / Classification

    eclass No.: 32160502
    Documents - Protocols - Downloads :
    Here you will find information and further literature. For further documents (certificates with additional lot numbers, safety data sheets in other languages, further product information) please contact Genaxxon biosience at: info@genaxxon.com or phone: +49 731 3608 123.


    Documents:

    Manuals