Test Catalog

Test Id : NAIFA

Antinuclear Antibodies, HEp-2 Substrate, IgG, Serum

Useful For
Suggests clinical disorders or settings where the test may be helpful

Evaluation of patients suspected of having systemic autoimmune rheumatic disease (ANA-associated rheumatic diseases or connective tissue disease) or organ-specific autoimmune diseases such as autoimmune liver diseases

Method Name
A short description of the method used to perform the test

Indirect Immunofluorescence

NY State Available
Indicates the status of NY State approval and if the test is orderable for NY State clients.


Reporting Name
Lists a shorter or abbreviated version of the Published Name for a test

Antinuclear Ab, HEp-2 Substrate, S

Lists additional common names for a test, as an aid in searching

ANA (Antinuclear Antibodies)

Antinuclear Antibodies (ANA), HEp-2 substrate, Serum

LE (Lupus Erythematosus) Prep

Antinuclear Antibody, Serum



FANA (Fluorescent Antinuclear Antibodies

Specimen Type
Describes the specimen type validated for testing


Specimen Required
Defines the optimal specimen required to perform the test and the preferred volume to complete testing

Container/Tube: Serum gel or red top

Specimen Volume: 0.5 mL


If not ordering electronically, complete, print, and send a General Request (T239) with the specimen.

Specimen Minimum Volume
Defines the amount of sample necessary to provide a clinically relevant result as determined by the Testing Laboratory

0.3 mL

Reject Due To
Identifies specimen types and conditions that may cause the specimen to be rejected

Gross hemolysis OK
Gross lipemia OK
Gross icterus OK

Specimen Stability Information
Provides a description of the temperatures required to transport a specimen to the performing laboratory, alternate acceptable temperatures are also included

Specimen Type Temperature Time Special Container
Serum Refrigerated (preferred) 21 days
Frozen 28 days

Useful For
Suggests clinical disorders or settings where the test may be helpful

Evaluation of patients suspected of having systemic autoimmune rheumatic disease (ANA-associated rheumatic diseases or connective tissue disease) or organ-specific autoimmune diseases such as autoimmune liver diseases

Clinical Information
Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test

Autoantibodies targeting antigens in the nuclear region in the HEp-2 cell line substrate using the indirect immunofluorescence assay (IFA) have traditionally been called antinuclear antibody (ANA). ANA is the commonly performed antibody test in the initial evaluation of patients with systemic autoimmune rheumatic diseases (also referred to as connective tissue disease). Classic ANA-associated rheumatic diseases include systemic lupus erythematosus (SLE), mixed connective tissue disease (MCTD), Sjogren syndrome (Sjs), and systemic sclerosis (SSc) including CREST (calcinosis, Raynaud phenomenon, esophageal dysmotility, sclerodactyly, telangiectasia) and inflammatory myopathies (IM) such as dermatomyositis (DM).(1-4) Testing for ANA may also be of diagnostic relevance in the differential evaluation of autoimmune liver diseases (ALD).(5-6)


The classical ANA patterns (antibodies targeting the nuclear region) include homogeneous, speckled, centromere, nuclear dots, and nucleolar. These patterns are routinely reported by most clinical laboratories. SLE patients and those with SSc, Sjs, IM (such as anti-synthetase syndrome and necrotizing autoimmune myopathy) or ALD have also been shown to have clinically significant antibodies that react with antigens in other cellular compartments such as the cytoplasm and structures associated mitosis or mitotic patterns with HEp-2 substrate (reviewed in 1-3). Based on the increasing recognition of these non-nuclear antigenic targets and their documented clinical relevance, the first International Consensus on ANA Patterns established a classification tree for ANA with alpha-numeric anti-cell (AC) code for each pattern with a recommendation for a change in terminology from antinuclear antibody to anticellular antibody.(2) These changes are relevant as in addition the nuclear patterns, the classification includes cytoplasmic and mitotic patterns with descriptions for their interpretation, associated antibody targets and clinical associations when available.(4)


The diagnosis of ANA-associated rheumatic diseases is usually based on a set of criteria of which the presence on anticellular antibody or specific associated antibodies may be components. Of all ANA-associated rheumatic diseases, the presence of anticellular antibody is considered mandatory entry criterion by the 2019 European League Against Rheumatism and the American College of Rheumatology classification criteria for SLE.(7) Since cytoplasmic staining patterns may be reported as "ANA negative" or as a comment with no quantitative or titer result, some patients with clinicopathological symptoms consistent with neuropsychiatric SLE would not qualify for entry based on where testing is performed.(8-10) This limitation may therefore exclude patients who may meet the clinical and other laboratory criteria for disease but are not reported as "ANA positive" due to the use of the current terminology. In an international inception cohort of newly diagnosed SLE patients, 6.2% were anticellular antibody-negative with 1.5% testing positive for isolated cytoplasmic or mitotic pattern.(11)


Although the anticellular antibody is a sensitive test, it lacks diagnostic specificity.(1-4) Therefore, positive results require confirmation with the use of specific ANA-associated antibody tests except for the centromere pattern which is very characteristic for patients with limited diffuse SSc. Confirmation of a positive anticellular antibody test result may be guided by HEp-2 IFA patterns and/or titer, patient’s clinical presentation or in some cases the patient’s demographic.(12)

Reference Values
Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.

<1:80 (Negative)

Provides information to assist in interpretation of the test results

Presence of anticellular antibody (formerly 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.


Anticellular 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.

Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances

Some patients without clinical evidence of systemic autoimmune rheumatic disease (SARD) maybe positive for anticellular antibody. This occurs at variable prevalence depending on the patient demographics. A positive result may also precede clinical manifestation of SARD or be associated with some viral or chronic infections, cancers, or use of certain medications. All results must be reported in the appropriate clinical context as the performance of the test can be variable.

Clinical Reference
Recommendations for in-depth reading of a clinical nature

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;73:17-23

2. Chan EK, Damoiseaux J, Gabriel Carballo O, 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;16:715-726

4. International Consensus on ANA Patterns: Nomenclature and Classification Tree. ICAP; 2021 Accessed August 13, 2021. 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;67:145-172

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

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;71:1400-1412

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;47:1768-1773

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;11:17

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;12:14

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

12. Nandjwada 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;43:1816-1824

Method Description
Describes how the test is performed and provides a method-specific reference

Antibodies to nuclear antigens in a human epithelial type 2 (HEp-2) cell line by an indirect immunofluorescent technique. Commercial slides prepared from HEp-2 cells are used as a substrate. IgG antibodies in serum specimens are detected after incubation of serum with the commercial slides by the addition of a fluorescein isothiocyante (FITC)-labeled antihuman-IgG reagent. All patient specimens are initially screened at 1:80.(Package insert: NOVA Lite  DAPI ANA. Inova Diagnostics; 05/2015)

PDF Report
Indicates whether the report includes an additional document with charts, images or other enriched information


Day(s) Performed
Outlines the days the test is performed. This field reflects the day that the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time before the test is performed. Some tests are listed as continuously performed, which means that assays are performed multiple times during the day.

Monday through Saturday

Report Available
The interval of time (receipt of sample at Mayo Clinic Laboratories to results available) taking into account standard setup days and weekends. The first day is the time that it typically takes for a result to be available. The last day is the time it might take, accounting for any necessary repeated testing.

2 days

Specimen Retention Time
Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded

14 days

Performing Laboratory Location
Indicates the location of the laboratory that performs the test


Several factors determine the fee charged to perform a test. Contact your U.S. or International Regional Manager for information about establishing a fee schedule or to learn more about resources to optimize test selection.

  • Authorized users can sign in to Test Prices for detailed fee information.
  • Clients without access to Test Prices can contact Customer Service 24 hours a day, seven days a week.
  • Prospective clients should contact their Regional Manager. For assistance, contact Customer Service.

Test Classification
Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR) product.

This test has been cleared, approved, or is exempt by the US Food and Drug Administration and is used per manufacturer's instructions. Performance characteristics were verified by Mayo Clinic in a manner consistent with CLIA requirements.

CPT Code Information
Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Clinic Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.

CPT codes are provided by the performing laboratory.


LOINC® Information
Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the order and results codes of this test. LOINC values are provided by the performing laboratory.

Test Id Test Order Name Order LOINC Value
NAIFA Antinuclear Ab, HEp-2 Substrate, S 59069-5
Result Id Test Result Name Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
ANAH Antinuclear Ab, HEp-2 Substrate, S 59069-5
1TANA ANA Titer: 33253-6
1PANA ANA Pattern: 49311-4
2TANA ANA Titer 2: 33253-6
2PANA ANA Pattern 2: 49311-4
CYTQL Cytoplasmic Pattern: 55171-3
LCOM Lab Comment: 77202-0

Test Setup Resources

Setup Files
Test setup information contains test file definition details to support order and result interfacing between Mayo Clinic Laboratories and your Laboratory Information System.

Excel | Pdf

Sample Reports
Normal and Abnormal sample reports are provided as references for report appearance.

Normal Reports | Abnormal Reports

SI Sample Reports
International System (SI) of Unit reports are provided for a limited number of tests. These reports are intended for international account use and are only available through MayoLINK accounts that have been defined to receive them.

SI Normal Reports | SI Abnormal Reports