Antinuclear Antibodies in Non-Rheumatic Diseases

Recommended by ACR and EULAR as the gold standard for ANA testing (Meroni and Schur 2010)

The most established method for ANA screening (Kumar et al. 2009; Pisetsky 2011)

Uses human epithelial (Hep-2) cells

Confirms nuclear antigens and fluorescence patterns using a fluorescence microscope (e.g., speckled, homogeneous, nuclear mixture, and cytoplasmic mixture patterns; Peng et al. 2014)

A negative IIF result is conclusive

All positive results must be confirmed by a second independent method (e.g., ELISA or ImmunoBlot) (Choi et al. 2020)

Determines staining patterns and titers concurrently (Agmon-Levin et al. 2014)

High sensitivity test

A time-consuming and labor-intensive test

A lack of adequate standardization (Damoiseaux et al. 2005; Agmon-Levin et al. 2014)

Poor specificity—false-positive results due to the non-specific binding of antibodies and the presence of closely related antigens sharing similar epitopes (Kątnik-Prastowska 2009)

Low positive predictive value

Operator-dependent test—high inter and intra-laboratory variabilities of pattern recognition resulted from the individual abilities of investigators (Infantino et al. 2017; Rigon et al. 2017)

Test based on antigen–antibody reaction

Automated methods (Shah and Maghsoudlou 2016)

Monospecific tests identifying various specific antigens in CTD such as dsDNA, SSA/Ro (52 kDA and 60 kDA), SSB/La, U1-RNP, Sm, centromere, Jo-1, Scl-70, Rib-P, fibrillarin, RNA Pol III, PM-Scl, PCNA and Mi-2 (Robier et al. 2015)

More specific and sensitive than IIF (Murdjeva et al. 2011; Aydin 2015; Kohl and Ascoli 2017)

Simple procedure

High specificity and sensitivity caused by an antigen–antibody reaction

High efficiency caused by lack of necessity of complicated sample pre-treatment

Safe and eco-friendly (radioactive substances and large amounts of organic solvents are not required)

Cost-effective assay (low-cost reagents)

Tedious/laborious assay procedure

Expensive preparation of antibodies, which requires specific culture cell media

The risk of antibodies inactivation caused by their labeling

High possibility of false positive/negative antigens detection resulting from cross-reactivity of secondary antibody (false results caused by insufficient blocking of immobilized antigen)

Antibody instability if not refrigerated transport and inadequate storage is performed (Konstantinou 2017)

Enables the detection of protein expression levels based on its molecular weight and immunoreactivity with a specific antibody (Butler et al. 2019)

Identifies antigen specificity (reactivity) on a membrane in distinct lines (Kroon et al. 2022)

Consists of a series of interrelated steps (Murdjeva et al. 2011)

More sensitive than DID and CIE

Particularly useful in detecting ANAs in non-rheumatic diseases (Pillai-Kastoori et al. 2020)

High sensitivity, specificity, ease, and rapidity of execution

Potential for automation

Precisely identifies antibody reactions against nuclear antigens (Lallier et al. 2019)

Expensive

Time consuming

Detects linear epitopes only

Minor variations in a series of interrelated steps can alter the quality of the blot and introduce errors (Pillai-Kastoori et al. 2020)

Reproducibility and trustworthiness (caused by non-precise test performance) (Butler et al. 2019; Kroon et al. 2022)

Evaluates antibody reactivity using beads with fluorescent signals and antigens

(Vignali 2000; Naides et al. 2020)

Concurrent antigen recognition

High sensitivity (Vignali 2000; Avaniss-Aghajani et al. 2007; Bonilla et al. 2007)

Economical multi-antigen testing

Has the potential for automation (Bonilla et al. 2007)

Lack of uniformity in testing may be a cause of misdirecting results (Naides et al. 2020)

Provides a single result at a time (Vignali 2000; Avaniss-Aghajani et al. 2007; Bonilla et al. 2007)

Based on the application of autoantigen array and flow technologies (Giavedoni 2005; Swana et al. 2019)

Is used in various immunoassay, genomic, and proteomic analyses (Giavedoni 2005; Satoh et al. 2015; Swana et al. 2019).

One serum may react with many autoantigens in a multiplexed ALBIA (Fritzler et al. 2006)

The ability to assess several antibody specificities simultaneously in a small serum sample

Reliable, accurate, cost-saving

Efficient

Has a rapid turnaround time

High sensitivity

Less dependence on highly skilled operators (Fritzler et al. 2006; Satoh et al. 2015; Swana et al. 2019).

N = 89

control: n = 100

20.2% (18 patients)

11 patients (23%)

n = 83

control: n = 83

33% with TB (mainly present anti-Scl-70)

20% in control group

ANAs titer: 1:1280 in pleural fluid

ANA IgG and IgM levels positive at 1:160 and 1:40, respectively

ANA titer inversely linked to schistosome infection

↑ ANA level after anti-helminthic treatment

↑ ANA prevalence in patients with the infection progression

6.7% in acute stage

23.3% in chronic stage

70.0% in late-stage

22% with AID

71% with CAH

Anti-ssDNA and anti-dsDNA in 60% of patients with ALD and CAH

NHL: n = 119

HL: n = 60

ANAs presence of patients with NHL without the risk of autoimmune disease development

n = 347

control: n = 213

ANAs occurrence targeting mitotic proteins

ANAs in 19% of NHL patients with particular subgroups, such as ANAs directed against mitotic proteins or associated mitotic proteins

ANA positive in 34% with leukemia

ANAs with a nucleolar pattern were linked to a two-fold increase in the risk of death compared to negative ANA results in leukemia

ANA positivity in 30 (13.9%) patients

The correlation of positive ANA and TP53 disruption improves accuracy in predicting OS

n = 120

control: n = 372

ANA detected in three EPF patients (2.5%)

n = 50

control n = 50

ANA positivity in 40% of patients

The most common patterns: homogenous or speckled

Rat esophagus is a preferred substrate for ANAs detection in LP patients, (ANA positive in 40.42% LP patients)

Monkey esophagus—ANA-positive in a 27.6% subjects, while Hep-2 cells and rat liver are unsuitable

ANA-positivity of 22%

ANA ≥1:160 in 53.5%

High ANAs titers are more prevalent in females (55.8%) than in males (44.15%)

10 patients

Ps: n = 101

PsA: n = 100

control: n = 50

25.7% Ps ANAs positive

15% PsA ANAs positive

n = 118

control: n = 50

↑ prevalence of ANAs in males (M: F = 57.1% vs. 42.9%)

n = 70

control: n = 20

27% of patients

Pediatric: n = 80

Pediatric: control n = 473

Adult: control n = 1125

Significantly higher prevalence (16.2%) of ANAs among pediatric diabetics

24% of subjects with CD (P < 0.001)

ANA positivity is associated with the presence of the HLA DQ2/DQ8 haplotypes (P < 0.001).

8.4% ANA positive

Showed distinct nuclear staining patterns: homogeneous (6), fine-speckled (7), and coarse-speckled (1)

GD: n = 256

Grave's orbitopathy n = 156

control: n = 78

ANA positive in 212 (80%) GD patients vs. ANA positive in no GO (91%)

n = 168

control: n = 75

ANAs in 35% of patients with ATD (anti-_ dsDNA and anti-Ro)

60% of patients have a speckled ANA pattern. (ANAs related to higher prevalence of ATG and ATPO in 30% of children with chronic lymphocytic thyroiditis without rheumatologic conditions

n = 124

control: 41

16.1% ANA

Positive ANAs (392.23 IU/mL; N = < 23) present in the course of antiphospholipid syndrome

AD co-exist with SLE