Test Catalog

Test Id : ANA2

Antinuclear Antibodies (ANA), Serum

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

Evaluating patients at-risk for antinuclear antibodies-associated systemic autoimmune rheumatic disease particularly systemic lupus erythematosus, Sjogren syndrome, and mixed connective tissue disease

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

Special Instructions
Library of PDFs including pertinent information and forms related to the test

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

Enzyme-Linked Immunosorbent Assay (ELISA)

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

Yes

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

Antinuclear Ab, S

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

ANA1

ANA2

Antinuclear Antibody, Serum

Hep-2

ANA (order test ANA2)

ANA (Antinuclear Antibodies)

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

LE (Lupus Erythematosus) Prep

ANA Screen

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

Specimen Type
Describes the specimen type validated for testing

Serum

Ordering Guidance

If suspicious of connective tissue disorder, see CTDC / Connective Tissue Disease Cascade, Serum.

 

If suspicious of autoimmune liver disease, see ALDG / Autoimmune Liver Disease Panel, Serum.

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

Container/Tube:

Preferred: Serum gel

Acceptable: Red top

Specimen Volume: 0.5 mL

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Forms

If not ordering electronically, complete, print, and send 1 of the following forms with the specimen:

-General Request (T239)

-Gastroenterology and Hepatology Client Test Request (T728)

-Renal Diagnostics Test Request (T830)

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

0.4 mL

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

Gross hemolysis Reject
Gross lipemia Reject
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 21 days

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

Evaluating patients at-risk for antinuclear antibodies-associated systemic autoimmune rheumatic disease particularly systemic lupus erythematosus, Sjogren syndrome, and mixed connective tissue disease

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

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

Measurement of antinuclear antibodies (ANA) in serum is the most commonly performed screening test for patients suspected of having a systemic autoimmune rheumatic disease (SARD), also referred to as connective tissue disease.(1) ANA occur in patients with various autoimmune diseases, both systemic and organ specific, but they are particularly common in SARDs, which  include systemic lupus erythematosus (SLE), discoid lupus erythematosus, drug-induced lupus erythematosus, mixed connective tissue disease (MCTD), Sjogren’s syndrome (SjS), systemic sclerosis, CREST syndrome (calcinosis, Raynaud’s phenomenon, esophageal dysmotility, sclerodactyly, telangiectasia), and idiopathic inflammatory myopathies.

 

ANA can be detected by different technologies, including indirect immunofluorescence assay (IFA) and solid phase assays, such as enzyme immunoassays and multiplex bead immunoassays. In a study performed in the Mayo Clinic Antibody Immunology Laboratory, no significant differences were demonstrated between ANA IFA and ANA enzyme-linked immunosorbent assay (ELISA) for a cohort of patients with connective tissue disease consisting predominantly of patients with SLE, SjS, and MCTD. Weakly positive ANA ELISA results were not a strong indicator of SARD in this laboratory cohort. The likelihood of finding an autoantibody to a specific extractable nuclear antigen including double-stranded DNA on a second-order testing increased directly with the level of ANA: 88% of sera that had detectable autoantibodies on second-order testing had an ANA level greater than 3.0 U.(2)

 

Overall, an ANA ELISA result of greater than or equal to 3.0 U was demonstrated as the optimal cutoff for CTDC / Connective Tissue Disease Cascade, Serum. This algorithm is intended to evaluate patients with common connective tissue diseases such as SLE, SjS, and MCTD.

 

See Connective Tissue Disease Cascade (CTDC)

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.

Negative: < or =1.0 U

Weakly positive: 1.1-2.9 U

Positive: 3.0-5.9 U

Strongly positive: > or =6.0 U

Reference values apply to all ages.

Interpretation
Provides information to assist in interpretation of the test results

A large number of healthy individuals have weakly-positive (1.1 - 2.9 U) antinuclear antibody (ANA) enzyme-linked immunosorbent assay (ELISA) results, many of which are likely to be clinical false-positive results; therefore, second-order testing of all positive ANA yields a very low percentage of positive results to extractable nuclear antigens including double-stranded (ds) DNA.(2)

 

Positive ANA results greater than 3.0 U are associated with the presence of detectable autoantibodies to specific extractable nuclear antigens (SM, SS-A, SS-B, Sm/RNP or RNP 68 and RNP A, Jo-1, Scl-70) including dsDNA.

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

Results for antinuclear antibodies (ANA) performed by enzyme-linked immunosorbent assay (ELISA) and traditional immunofluorescence assay (IFA) in the Mayo Clinic Antibody Immunology Laboratory show both methods to substantially equivalent for the evaluation of common connective tissue diseases (CTD; systemic lupus erythematosus, Sjogren syndrome , and mixed connective tissue disease).(2) Negative results do not rule out the diagnosis of other CTD, such as systemic sclerosis and inflammatory myopathies, and overlap syndromes associated with complex antigens optimally detected in with the ANA IFA and associated with nucleolar, certain cytoplasmic and/or speckled patterns. For autoimmune hepatitis and juvenile idiopathic arthritis, ANA IFA remains the optimal testing method as the target autoantigens are largely unknown and not included in the ELISA or other solid-phase immunoassays.(3-6)

 

ANA may also be detectable following viral illnesses, in chronic infections, or in patients treated with many different medications.

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 Jan;73(1):17-23. doi: 10.1136/annrheumdis-2013-203863

2. Deng X, Peters B, Ettore MW, et al: Utility of antinuclear antibody screening by various methods in a clinical laboratory patient cohort: J Appl Lab Med. 2016 Jul 1;1(1):36-46. doi: 10.1373/jalm.2016.020172

3. Sparchez M, Delean D, Samasca G, Miu N, Sparchez Z: Antinuclear antibody screening by ELISA and IF techniques: discrepant results in juvenile idiopathic arthritis but consistency in childhood systemic lupus erythematous. Clin Rheumatol. 2014 May;33(5):643-647. doi: 10.1007/s10067-014-2529-y

4. Bossuyt X, De Langhe E, Borghi MO, Meroni PL: Understanding and interpreting antinuclear antibody tests in systemic rheumatic diseases. Nat Rev Rheumatol. 2020 Dec;16(12):715-726. doi: 10.1038/s41584-020-00522-w

5. Bossuyt X, Claessens J, De Langhe E, et al: Antinuclear antibodies by indirect immunofluorescence and solid phase assays. Ann Rheum Dis. 2020 Jun;79(6):e65. doi: 10.1136/annrheumdis-2019-215443

6. Alsaed OS, Alamlih LI, Al-Radideh O, Chandra P, Alemadi S, Al-Allaf AW. Clinical utility of ANA-ELISA vs ANA-immunofluorescence in connective tissue diseases. Sci Rep. 2021 Apr 15;11(1):8229. doi: 10.1038/s41598-021-87366-w

Special Instructions
Library of PDFs including pertinent information and forms related to the test

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

The method used to detect antinuclear antibody (ANA) is enzyme-linked immunosorbent assay (ELISA). A HEp-2 lysate supplemented with specific purified antigens (double-stranded deoxyribonucleic acid [dsDNA], histone, SS-A [Ro], SS-B [La] Smith, sm/RNP, Scl-70, Jo-1, and centromere B antigen) are coated onto microtiter plate wells. A dilution of patient serum is added to the well and incubated. After washing to remove unbound serum protein, an enzyme-conjugated antihuman-IgG antibody is added to detect human IgG bound to the microtiter plate well. After incubation and washing to remove unbound conjugate, a substrate to the enzyme is added to the well. After incubation, the enzyme substrate reaction is stopped. The complete assay is measured on a spectrophotometer plate reader. The optical density measured is proportional to the antibody present in the patient serum. Testing is performed on the Agility instrument by Dynex.(Package insert: ELISA kits. Bio-Rad Laboratories; 07/14)

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

No

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.

1 day

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

Rochester

Fees
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.

86038

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