Test Code RFLXP Antinuclear Antibodies, HEp-2, Reflex Panel

Specimen Required

Only orderable as a reflex. For more information see RAIFA / Antinuclear Antibodies, HEp-2 Substrate, IgG, with Reflex, Serum.

Supplies: Sarstedt Aliquot Tube, 5 mL (T914)

Collection Container/Tube:

Preferred: Serum gel

Acceptable: Red top

Submission Container/Tube: Plastic vial

Specimen Volume: 0.7 mL Serum

Collection Instructions: Centrifuge and aliquot serum into a plastic vial.

Useful For

Confirmatory testing after a positive homogenous, speckled, or dense fine speckled pattern identified in the human epithelial type 2 antinuclear antibody indirect immunofluorescence assay

Profile Information

Test ID	Reporting Name	Available Separately	Always Performed
ADNA1	dsDNA Ab, IgG, S	Yes	Yes
RNP	RNP Ab, IgG, S	Yes	Yes
SCL70	Scl 70 Ab, IgG, S	Yes	Yes
SM	Sm Ab, IgG, S	Yes	Yes
SSA	SS-A/Ro Ab, IgG, S	Yes	Yes
SSB	SS-B/La Ab, IgG, S	Yes	Yes
IM_04	Antinuclear Ab,HEp-2,reflex Comment	No	No

Method Name

Only orderable as a reflex. For more information see RAIFA / Antinuclear Antibodies, HEp-2 Substrate, IgG, with Reflex, Serum.

ADNA1: Enzyme-Linked Immunosorbent Assay (ELISA)

RNP, SCL70, SM, SSA, SSB: Multiplex Flow Immunoassay

IM_04: Technical Interpretation

Reporting Name

ANA Ab HEp-2 Reflex Panel

Specimen Type

Serum

Specimen Minimum Volume

Serum: 0.5 mL

Specimen Stability Information

Specimen Type	Temperature	Time
Serum	Refrigerated (preferred)	21 days
	Frozen	21 days

Reference Values

Only orderable as a reflex. For more information see RAIFA / Antinuclear Antibodies, HEp-2 Substrate, IgG, with Reflex, Serum.

DOUBLE-STRANDED DNA (dsDNA) ANTIBODIES, IGG, SERUM

<100 IU/mL (negative)

≥100 IU/mL (positive)

Negative is considered normal.

Reference values apply to all ages.

RNP ANTIBODIES, IGG, SERUM

<1.0 U (negative)

≥1.0 U (positive)

Reference values apply to all ages.

SCL 70 ANTIBODIES, IGG, SERUM

<1.0 U (negative)

≥1.0 U (positive)

Reference values apply to all ages.

SMITH (Sm) ANTIBODIES, IGG, SERUM

<1.0 U (negative)

≥1.0 U (positive)

Reference values apply to all ages.

SS-A/RO ANTIBODIES, IGG, SERUM

<1.0 U (negative)

≥1.0 U (positive)

Reference values apply to all ages.

SS-B/LA ANTIBODIES, IGG, SERUM

<1.0 U (negative)

≥1.0 U (positive)

Reference values apply to all ages.

Interpretation

Presence of anti-cellular antibody (also known as antinuclear antibody) is a feature of systemic autoimmune rheumatic diseases such as systemic lupus erythematosus, mixed connective tissue disease, Sjogren syndrome, and systemic sclerosis and some inflammatory myopathies (dermatomyositis, anti-synthetase syndrome and necrotizing autoimmune myopathy). It may also be of diagnostic relevance in patients with autoimmune liver diseases.

Patients' sera are screened at 1:80. The following nuclear patterns and their titers are reported: centromere, homogeneous, nuclear dots, nucleolar, speckled, fine dense speckled (also referred to as DFS70), and proliferating cell nuclear antigen (PCNA). If observed, the following cytoplasmic patterns are reported: reticular/AMA (antimitochondrial antibody), cytoplasmic speckled, fibrillar, polar/Golgi-like, or rods and rings. The spindle fiber and centrosome mitotic patterns are also reported if observed. Reported patterns may help guide differential diagnosis, although they may not be specific for individual antibodies or diseases. Negative results do not necessarily rule out systemic autoimmune rheumatic disease.

The anti-cellular antibody test lacks diagnostic specificity and is associated with some cancers, infectious, and inflammatory conditions, with variable prevalence in healthy individuals. The lack of diagnostic specificity requires confirmation of positive results using associated antibody tests such as those targeting extractable nuclear antigens.

Clinical Reference

1. Agmon-Levin N, Damoiseaux J, Kallenberg C, et al. International recommendations for the assessment of autoantibodies to cellular antigens referred to as anti-nuclear antibodies. Ann Rheum Dis. 2014;73(1):17-23

2. Chan EK, Damoiseaux J, Carballo OG, et al. Report of the First International Consensus on Standardized Nomenclature of Antinuclear Antibody HEp-2 Cell Patterns 2014-2015. Front Immunol. 2015;6:412

3. Bossuyt X, De Langhe E, Borghi MO, Meroni PL. Understanding and interpreting antinuclear antibody tests in systemic rheumatic diseases. Nat Rev Rheumatol. 2020;16(12):715-726

4. International Consensus on ANA Patterns. Nomenclature and Classification Tree. ICAP; 2021 Accessed April 11, 2025. Available at www.anapatterns.org/trees.php

5. European Association for the Study of the Liver. EASL Clinical Practice Guidelines: The diagnosis and management of patients with primary biliary cholangitis. J Hepatol. 2017;67(1):145-172

6. Younossi ZM, Bernstein D, Shiffman ML, et al. Diagnosis and management of primary biliary cholangitis. Am J Gastroenterol. 2019;114(1):48-63

7. Aringer M, Costenbader K, Daikh D, et al. 2019 European League Against Rheumatism/American College of Rheumatology classification criteria for systemic lupus erythematosus. Arthritis Rheumatol. 2019;71(9):1400-1412

8. Naides SJ, Genzen JR, Abel G, Bashleben C, Ansari MQ. Antinuclear antibodies testing method variability: A survey of participants in the College of American Pathologists' Proficiency Testing Program. J Rheumatol. 2020;47(12):1768-1773

9. Van Hoovels L, Broeders S, Chan EKL, et al. Current laboratory and clinical practices in reporting and interpreting anti-nuclear antibody indirect immunofluorescence (ANA IIF) patterns: results of an international survey. Auto Immun Highlights. 2020;11(1):17

10. Tebo AE, Schmidt RL, Kadkhoda K, et al. The antinuclear antibody HEp-2 indirect immunofluorescence assay: a survey of laboratory performance, pattern recognition and interpretation. Auto Immun Highlights. 2021;12(1):4

11. Choi MY, Clarke AE, St Pierre Y, et al. Antinuclear antibody-negative systemic lupus erythematosus in an international inception cohort. Arthritis Care Res (Hoboken). 2019;71(7):893-902

12. Nandiwada SL, Peterson LK, Mayes MD, et al. Ethnic differences in autoantibody diversity and hierarchy: More clues from a US cohort of patients with systemic sclerosis. J Rheumatol. 2016;43(10):1816-1824

13. Silva MJ, Dellavance A, Baldo DC, et al. Interkit Reproducibility of the Indirect Immunofluorescence Assay on HEp-2 Cells Depends on the Immunofluorescence Reactivity Intensity and Pattern. Front Immunol. 2022;12:798322

Day(s) Performed

Monday through Saturday

Report Available

3 to 4 days

Performing Laboratory

Mayo Clinic Laboratories in Rochester

Test Classification

Not Applicable

CPT Code Information

86225

86235 x5

LOINC Code Information

Test ID	Test Order Name	Order LOINC Value
RFLXP	ANA Ab HEp-2 Reflex Panel	97564-9

Result ID	Test Result Name	Result LOINC Value
ADNA1	dsDNA Ab, IgG, S	33799-8
IM_04	Antinuclear Ab,HEp-2,reflex Comment	77202-0
RNP	RNP Ab, IgG, S	29958-6
SCL70	Scl 70 Ab, IgG, S	47322-3
SM	Sm Ab, IgG, S	18323-6
SSA	SS-A/Ro Ab, IgG, S	33610-7
SSB	SS-B/La Ab, IgG, S	33613-1