Autoimmune Diseases and Antinuclear Antibody Testing: Methods and Staining Patterns
Continuing Education Credits
This course concentrates and expands upon both the diagnosis and laboratory testing for autoimmune diseases, as well as on the interpretation of antinuclear antibody (ANA) staining patterns. Laboratory methods for detection and identification of ANAs are complex. Proper training is required to correctly interpret results at both the laboratory and clinical level. This educational program provides a comprehensive review of the methods, interpretation, and clinical significance of ANA testing. The material presented will describe various autoimmune diseases with emphasis on diagnosis.
Objectives
- Discuss the autoimmune reaction and the various types of autoimmune diseases.
- Describe the signs, symptoms, and causes of autoimmune diseases.
- Describe the laboratory tests and exams used to diagnose autoimmune diseases.
- Define the antinuclear antibody test (ANA) and explain the differences in ANA methods.
- Distinguish positive from negative samples.
- Identify and classify ANA patterns.
- Discuss the clinical significance of ANAs.
Course Outline
- Copyright and Disclosure
- Copyright
- Disclosure
- Overview of Autoimmune Diseases
- Autoimmune Disease Definition and Overview
- Immune Mechanisms Preventing Autoimmunity
- Loss of Self Tolerance
- Initiation of Autoimmune Disease
- Which of the following statements is true with regard to autoimmune diseases?
- True or False: Gender and genetic predisposition are factors in the development of an autoimmune reaction. Another factor is the occurrence of a triggering event that leads to a breakdown of tolerance.
- Table 1: Common Autoimmune Diseases
- Systemic Autoimmune Rheumatic Diseases
- Systemic Autoimmune Rheumatic Diseases, continued
- Which of these is not considered one of the systemic autoimmune rheumatic diseases (SARDs)?
- In which autoimmune disorder (and a form of SARD) does the body's immune system mistakenly attack the tissue in glands that produce moisture, such as tear and salivary glands?
- Signs, Symptoms, Causes, and Treatment of Autoimmune Diseases
- Diagnosis of Autoimmune Diseases
- Laboratory Tests: Inflammatory Markers
- Laboratory Tests: Autoantibodies
- The initial laboratory evaluation for autoimmune diseases often involves the following tests, except:
- Introduction to ANA Testing
- Antinuclear Antibody Test
- Antinuclear Antibody Test: Background, History, and Overview
- Cell Morphology (HEp-2 or HEp-2000 Cell)
- Stages of Mitosis
- Which statement is false when describing the ANA test?
- Which two stages of cell division are the most important for reading ANAs?
- Match each letter in this image of a tissue cell used as a substrate in ANA testing to the structure it indicates.
- ANA Testing Methods
- Indirect Immunofluorescence Assay
- Fluorescent ANA Testing
- Colorzyme®
- Which statement is false when describing indirect immunofluorescence assays (IFAs)?
- Which statement accurately describes the Colorzyme® system?
- True or False: Depending on the laboratory and specific method used, an ANA titer is typically considered negative when the titer is equal to or less than 1:80.
- Enzyme-Linked Immunosorbent Assay
- Other Methods for ANA Testing
- Conditions Associated with a Positive ANA Test
- About 95% of those individuals with a particular condition have a positive ANA test. What is the most common condition associated with a positive ANA test?
- ANA Pattern Interpretation
- Reading ANA Patterns Using a HEp-2 or HEp-2000® Substrate
- For the ANA test to be positive, what must be present in the nucleus of the interphase cell?
- ANA Testing Scheme
- Why are ANA positive samples titered?
- Follow-up Testing
- Table 2: Antigen Chart
- ANA Patterns
- ANA Patterns
- Negative
- Homogeneous (AC-1)
- Speckled (AC-5)
- Nucleolar (AC-9)
- Centromere (AC-3)
- SS-A/Ro Pattern
- Proliferating Cell Nuclear Antigen (AC-13)
- Nuclear Membrane (AC-11)
- The image on the right represents the result of a fluorescent antinuclear antibody (ANA) test. What pattern should be reported?Note: (a) points to the nuclei of several interphase cells, the primary consideration for discerning the ANA pattern, and (b) indicates a metaphase mitotic cell. Observing the chromosomal area and cytoplasm of the metaphase cell may assist in identifying the ANA pattern.
- Unusual or Atypical Speckled Patterns
- Unusual or Atypical Speckled Patterns: Overview
- Description of Unusual or Atypical Speckled Patterns
- Description of Unusual or Atypical Speckled Patterns, continued
- True or False: When identifying the midbody pattern, only cells in interphase and metaphase are useful.
- Mixed ANA Pattterns
- Mixed ANA Patterns: Overview
- Homogeneous and Speckled
- Homogeneous and Centromere
- Homogeneous and SS-A/Ro (on HEp-2000®)
- Speckled and SS-A/Ro Pattern
- Scl-70 Pattern
- Other Mixed Patterns
- What is the most common combination of ANA patterns?
- Cytoplasmic Patterns
- Cytoplasmic ANA Patterns: Overview
- Common Cytoplasmic Staining Patterns
- Common Cytoplasmic Staining Patterns, continued
- True or False: Cytoplasmic patterns are interesting to look at, but have no clinical significance and should not be reported.
- References
- References
Additional Information
Level of Instruction: Intermediate
Intended Audience: Clinical laboratory professionals, including medical laboratory scientists and medical laboratory technicians. This course is also appropriate for medical laboratory students, pathologists, clinicians, and rheumatologists.
Author Information:
Robert R. Boyes is the former Senior Vice President and General Manager for Immuno Concepts in Sacramento, California. He has a degree in Medical Technology. In his career as a clinical laboratory professional, he has worked in hospital laboratories, specialty clinics, and the corporate setting. Mr. Boyes has conducted numerous research presentations and has authored or co-authored several publications related to antinuclear antibodies.
David J. Moffa, PhD, BCLD, has over 30 years of experience in the healthcare industry as an executive manager, clinical laboratory director, and medical laboratory scientist. He is currently a technical consultant for Kentmere Healthcare, Wilmington, DE, and until his retirement, was the Regional Director for LabCorp, Inc. He holds a PhD in medical biochemistry from the School of Medicine, West Virginia University.
Reviewer Information:
Kaitlin Faughnan, MS, MLS(ASCP)CM, is currently a faculty member of SUNY Broome Community College, where she teaches all subjects of MLS to both Medical Laboratory Technician and Histotechnician students. She is involved in curriculum development and revision, as well as researching updates to the field of laboratory science to stay on the cutting edge of the latest information and technology. She received both her Bachelor of Science and Master of Science in Clinical Laboratory Science. She holds New York state licensure and previously worked as a generalist in a hospital.
Laurie Bjerklie, MA, MLS(ASCP)CM, is
a Lead Education Developer. She earned a B.S. in Medical Laboratory Science
from the University of North Dakota and an M.A. in Curriculum and Instruction
from Saint Xavier University. She has over 15 years of experience in higher
education and has held program director and faculty positions in both MLT and
MLS programs.
Course Description: This course concentrates and expands upon the diagnosis and laboratory testing for autoimmune diseases, as well as the interpretation of antinuclear antibody (ANA) staining patterns. Laboratory methods for the detection and identification of ANAs are complex. Proper training is required to correctly interpret results at both the laboratory and clinical levels. This educational program provides a comprehensive review of the methods, interpretation, and clinical significance of ANA testing.