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HLA -B27 Test: Introduction, Principle, Procedure, Result-Interpretation, Clinical Significance, and Keynotes

Introduction

HLA-B27 (Human Leukocyte Antigen B27) is a specific genetic variant of the HLA-B gene located on chromosome 6. While HLA proteins normally help the immune system distinguish between the body’s own cells and foreign invaders, the B27 variant is a well-known risk factor for a group of autoimmune disorders known as seronegative spondyloarthropathies.

Principle

The test operates on the principle of detecting either the HLA-B27 protein on the surface of lymphocytes or the HLA-B27 gene in the individual’s DNA.

  • Protein Detection: Uses specific antibodies that bind to the HLA-B27 antigen if it is present on the white blood cell surface.
  • Genetic Detection: Uses molecular techniques to identify the specific DNA sequence corresponding to the HLA-B27 allele.

Procedure

Testing typically involves a standard blood draw (venipuncture). Common laboratory methods include:

  • Flow Cytometry: A rapid, high-throughput method where blood cells are tagged with fluorescent antibodies and passed through a laser to detect the B27 protein.
  • PCR (Polymerase Chain Reaction): The current “gold standard” molecular method that amplifies and identifies the HLA-B27 gene directly from leukocyte DNA.
  • Microlymphocytotoxicity (MLCT): A traditional serological method where lymphocytes are incubated with B27-specific antisera and complement; cell death indicates a positive result.

Result Interpretation

Results are generally reported as Positive or Negative.

  • Positive Result: Indicates the presence of the HLA-B27 antigen/gene. This signifies a higher statistical risk for associated autoimmune disorders, but is not a diagnosis on its own.
Fig. Interpretation of HLA-B27 Result
  • Negative Result: Indicates the antigen was not detected. While it significantly reduces the likelihood of HLA-B27-related diseases, it does not completely rule them out, as some patients develop these conditions without the marker.

Clinical Significance

The test is primarily used to support a diagnosis of:

  • Ankylosing Spondylitis (AS): Strongest association; ~90% of AS patients are HLA-B27 positive.
  • Reactive Arthritis (Reiter’s Syndrome): Often triggered by an infection.
  • Acute Anterior Uveitis: Recurring inflammation of the eye.
  • Psoriatic Arthritis & Inflammatory Bowel Disease (IBD)-associated Arthritis: Other conditions within the seronegative spondyloarthropathy family.

Keynotes

  • Not Diagnostic Alone: Most people with the HLA-B27 gene never develop an autoimmune disease.
  • Demographics: Prevalence varies by ethnicity; for example, it is more common in Northern European populations and less common in African American populations.
  • Lifelong Marker: Once a person tests positive, they will always be positive; the test does not need to be repeated.
  • Cross-Reactivity: Some tests (like flow cytometry) may occasionally show false positives due to cross-reactivity with similar antigens like HLA-B7.

Further Readings

  1. https://healthservice.hse.ie/documents/8275/Testing_for_HLA_B27.pdf
  2. https://pmc.ncbi.nlm.nih.gov/articles/PMC6817030/
  3. https://www.yashodahospitals.com/diagnostics/hla-b27-test/
  4. https://www.apollohospitals.com/diagnostics-investigations/hla-b27-test
  5. https://www.metropolisindia.com/blog/health-test/hla-b27-test-overview-procedure-and-result
  6. https://www.carehospitals.com/diagnostics/hla-b27-test
  7. https://www.ucsfhealth.org/care/medical-tests/hla-b27-antigen
  8. https://pubmed.ncbi.nlm.nih.gov/10932047/
  9. https://onlinelibrary.wiley.com/doi/full/10.1002/cyto.b.22102
  10. https://www.researchgate.net/publication/14314330_HLA-B27_determination_using_serological_methods_A_comparison_of_enzyme_immunoassay_and_a_microlymphocytotoxic_test_with_flow_cytometry_and_a_molecular_biological_assay
  11. https://labtestsonline.org.uk/tests/hla-b27
  12. https://www.ucsfbenioffchildrens.org/medical-tests/hla-b27-antigen
  13. https://punerheumatologist.com/blogs/hla-b27-test-ankylosing-spondylitis-facts/
  14. https://redcliffelabs.com/hla-b27-flow-cytometry
  15. https://www.ganeshdiagnostic.com/blog/what-is-hla-b27-pcr-test
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