ANA Screen,IFA, with Reflex to Titer and Pattern

Test Code

CPT Codes

If ANA Screen, IFA is positive, then ANA Titer and Pattern will be performed at an additional charge (CPT code(s): 86039).

Preferred Specimen
1 mL serum

Minimum Volume
0.5 mL

Transport Container
Transport tube

Transport Temperature
Room temperature

Specimen Stability
Room temperature: 4 days
Refrigerated: 7 days
Frozen: 30 days

Reject Criteria (Eg, hemolysis? Lipemia? Thaw/Other?)
Gross hemolysis • Grossly lipemic • Microbial contamination may interfere

Immunofluorescence Assay (IFA)

Setup Schedule

Report Available
1-4 days

Reference Range
ANA Screen Negative
  ANA Titer  
  <1:40 Negative
  1:40-1:80 Low antibody level
  >1:80 Elevated antibody level

Clinical Significance

This immunofluorescence assay (IFA) is often ordered as part of an initial diagnostic evaluation of individuals with clinical suspicion of autoimmune diseases associated with antinuclear antibodies (ANAs). The American College of Rheumatology (ACR) recommends IFA on human epithelial type 2 (HEp-2) cells as the gold standard method for ANA testing because of its overall high sensitivity [1].

ANAs are associated with several autoimmune diseases, such as systemic lupus erythematosus, systemic sclerosis, mixed connective tissue disease, polymyositis, primary biliary cholangitis, rheumatoid arthritis, juvenile rheumatoid arthritis, Sjogren syndrome, and autoimmune hepatitis. The laboratory evaluation for individuals with clinical suspicion of these autoimmune diseases often begins with an ANA screen.

Knowing the ANA titer can be helpful in interpreting positive ANA results. A titer of at least 1:40 is considered positive, but low-positive titers are not uncommon in healthy individuals. Higher titers are generally associated with greater likelihood of autoimmune disease [2]. When results are positive, various fluorescent staining patterns observed in the nucleus or the cytoplasm can aid in the differential diagnosis and guide selection of further testing for specific autoantibodies. The International Consensus on Antinuclear Antibody Pattern provides guidance on interpretation and reporting of IFA staining patterns with HEp-2 cells [3].

Individuals with negative results on the ANA IFA usually also have negative results on specific ANAs. Therefore, subserology testing is generally not recommended in individuals without positive ANA IFA results and clinical suspicion of relevant autoimmune disease [4]. However, Jo-1 antibody may be detected in ANA IFA-negative patients with some types of myositis, and SSA antibody may be detected in some ANA IFA-negative patients with lupus or Sjogren syndrome [4].

The results of this test should be interpreted in the context of pertinent clinical and family history and physical examination findings.

1. Methodology of testing for antinuclear antibodies (position statement). 2009. American College of Rheumatology. Updated December 2019. Accessed May 15, 2023.
2. Tozzoli R, et al. Am J Clin Pathol. 2002;117(2):316-324.
3. Chan EK, et al. Front Immunol. 2015;6:412.
4. Yazdany J, et al. Arthritis Care Res (Hoboken). 2013;65(3):329-339.

Performing Laboratory
Quest Diagnostics-Lenexa
10101 Renner Blvd
Lenexa , KS 66219-9752

The CPT Codes provided in this document are based on AMA guidelines and are for informational purposes only. CPT coding is the sole responsibility of the billing party. Please direct any questions regarding coding to the payor being billed. Any Profile/panel component may be ordered separately. Reflex tests are performed at an additional charge.