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The page below is a sample from the LabCE course Autoimmune Diseases and Antinuclear Antibody Testing: Methods and Staining Patterns. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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SSA/Ro Pattern

This is an example of the SSA/Ro ANA pattern using the HEp-2000® substrate. (This hyperexpressing pattern is not seen on standard HEp-2 substrates.)

As stated earlier, the HEp-2000® substrate utilizes genetically engineered HEp-2 cells with increased expression of SS-A/Ro antigen in the cells.

When the patient sample contains autoantibodies to the SS-A/Ro antigen, approximately 90% of the time the sample will produce a distinctive pattern with 10 -15% of the hyperexpressing cells showing strong speckled staining, frequently with strong nucleolar staining (a). The metaphase mitotic cells are negative (b). The remaining cells demonstrate weak speckled and nucleolar staining (c).

The presence of anti-SSA/Ro antibodies is confirmed by the unique staining pattern. Follow-up testing for antibodies to other extractable nuclear antigens (ENAs) is recommended. These antibodies are seen in patients with SLE, Sjögren's syndrome and low frequency in other diseases.

This pattern is reported as ANA positive, SSA/Ro pattern, anti-SSA/Ro antibodies present. Most labs will titer these samples. However, because the significance is that the anti-SSA antibodies are present regardless of the titer endpoint, some labs do not titer the SSA/Ro pattern.

SS-A/Ro (HEp-2000® only) Interphase:

Seen with about 89% of samples containing anti-SS-A/Ro antibodies

Hyperexpressing interphase cells

  • 10-15% hyperexpress the SS-A/Ro antigen
  • Stronger nucleolar staining
  • Stronger speckled staining

Non-hyperexpressing interphase cells

  • May or may not stain

SS-A/Ro (HEp-2000® only) Metaphase:

Metaphase mitotic cells

  • No staining in the chromosome region
  • Region outside of the chromosome area will stain with variable intensity
Some mitotics may also demonstrate brighter staining