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Kim, Ihm, Sin, Ihm, and Sim: Detection of Anti-ENA and anti-dsDNA Antibodies Using Line Immunoassay in Systemic Autoimmune Diseases
Original Article

Korean J Leg Med 2008;28(5):353-361.

Published online: 14 February 2008

DOI: https://doi.org/10.3343/kjlm.2008.28.5.353

Detection of Anti-ENA and anti-dsDNA Antibodies Using Line Immunoassay in Systemic Autoimmune Diseases

Ji Myung Kim, M.D.1, Chun Hwa Ihm, M.D.1, Dong Hyuk Sin, M.D.2, Mi Kyung Ihm, M.D.2, Seung Chul Sim, M.D.2

1Department of Laboratory Medicine, Eulji University Hospital, Daejeon, Korea

2Department of Rheumatology, Eulji University Hospital, Daejeon, Korea

Received 18 February 200830 June 2008       Accepted 21 July 2008

Copyright © 2008 by the Korean Society for Legal Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

Detection of antibodies to extractable nuclear antigens (ENAs) and dsDNA is needed for the diagnosis of and predicting prognosis in systemic autoimmune diseases. Recently introduced line immunoassay (LIA) has the advantage of detecting several autoantibodies simultaneously, and we evaluated its usefulness in the diagnosis of autoimmune diseases in comparison with enzyme-linked immunosorbent assay (ELISA).

Methods

Samples were collected from 437 patients referred by rheumatologists. FANA (fluorescent antinuclear antibody) test and LIA for the detection of 13 different autoantibodies, including 6 ENAs and dsDNA were performed. LIA-positive samples for ENA or dsDNA antibodies were further tested with ELISA. Final diagnosis was made by rheumatologists according to the diagnostic criteria. Agreement of results between LIA and ELISA was analyzed in 53 selected patients with systemic autoimmune diseases.

Results

The LIA detected antibodies to ENA and dsDNA in 118 and 22 patients, respectively, and ELISA detected 70.3% (83/118) and 45.5% (10/22) of LIA positive samples. Especially, 60.2% (71/118) of patients with positive ENA antibody on LIA was diagnosed as systemic autoimmune diseases. Patients having strong FANA titer and homogenous/speckled pattern showed higher prevalence of autoantibodies, but a small proportion of FANA negative patients also showed positive reactivity (LIA 10.8%, ELISA 5.2%). LIA showed a good agreement with ELISA for the anti-ENA antibodies (≥80%), and a lower agreement for the anti-dsDNA antibody (67.9%).

Conclusions

LIA detecting several autoantibodies simultaneously might replace ELISA for anti-ENA antibodies, but not for anti-dsDNA antibodies. When LIA is performed considering clinical manifestations and FANA, it could contribute to the diagnosis of systemic autoimmune disease.

Keywords: Autoimmune disease, Anti-ENA antibody, Anti-dsDNA antibody, Line immunoassay

REFERENCES

1.Reisner BS., DiBlasi J., Goel N. Comparison of an enzyme immunoassay to an indirect fluorescent immunoassay for the detection of antinuclear antibodies. Am J Clin Pathol. 1999. 111:503–6.
crossref
2.Fenger M., Wiik A., Hoier-Madsen M., Lykkegaard JJ., Rozenfeld T., Hansen MS, et al. Detection of antinuclear antibodies by solid-phase immunoassays and immunofluorescence analysis. Clin Chem. 2004. 50:2141–7.
crossref
3.Homburger HA. Cascade testing for autoantibodies in connective tissue disease. Mayo Clin Proc. 1995. 70:183–4.
4.Hoffman IE., Peene I., Veys EM., De Keyser F. Detection of specific antinuclear reactivities in patients with negative anti-nuclear antibody immunofluorescence screening tests. Clin Chem. 2002. 48:2171–6.
crossref
5.Bossuyt X., Luyckx A. Antibodies to extractable nuclear antigens in antinuclear antibody-negative samples. Clin Chem. 2005. 51:2426–7.
crossref
6.Delpech A., Gilbert D., Daliphard S., Le Loet X., Godin M., Tron F. Antibodies to Sm, RNP and SSB detected by solid-phase ELISAs using recombinant antigens: a comparison study with counter immunoelectrophoresis and immunoblotting. J Clin Lab Anal. 1993. 7:197–202.
7.Lopez-Longo FJ., Rodriguez-Mahou M., Escalona-Monge M., Gonzalez CM., Monteagudo I., Carreno-Perez L. Simultaneous identification of various antinuclear antibodies using an automated multi-parameter line immunoassay system. Lupus. 2003. 12:623–9.
crossref
8.Vitali C., Bombardieri S., Jonsson R., Moutsopoulos HM., Alexander EL., Carsons SE, et al. Classification criteria for Sjogren's syndrome: a revised version of the European criteria proposed by the American-European Consensus Group. Ann Rheum Dis. 2002. 61:554–8.
crossref
9.Hu PQ., Fertig N., Medsger TA Jr., Wright TM. Correlation of serum anti-DNA topoisomerase I antibody levels with disease severity and activity in systemic sclerosis. Arthritis Rheum. 2003. 48:1363–73.
crossref
10.Damoiseaux J., Boesten K., Giesen J., Austen J., Tervaert JW. Evaluation of a novel line-blot immunoassay for the detection of antibodies to extractable nuclear antigens. Ann N Y Acad Sci. 2005. 1050:340–7.
crossref
11.Rouquette AM., Desgruelles C., Laroche P. Evaluation of the new multiplexed immunoassay, FIDIS, for simultaneous quantitative determination of antinuclear antibodies and comparison with conventional methods. Am J Clin Pathol. 2003. 120:676–81.
crossref
12.Villalta D., Bizzaro N., Tonutti E., Visentin D., Manoni F., Piazza A, et al. Detection of anti-ENA autoantibodies in patients with systemic connective tissue disease. Analytical variability and diagnostic sensitivity of 4 methods. Recenti Prog Med. 1999. 90:579–84.
13.Damoiseaux JG., Tervaert JW. From ANA to ENA: how to proceed? Autoimmun Rev. 2006. 5:10–7.
crossref
14.Arbuckle MR., McClain MT., Rubertone MV., Scofield RH., Dennis GJ., James JA, et al. Development of autoantibodies before the clinical onset of systemic lupus eythematosus. N Eng J Med. 2003. 349:1526–33.
15.Sanchez-Guerrero J., Lew RA., Fossel AH., Schur PH. Utility of anti-Sm, anti-RNP, anti-Ro/SS-A, and anti-La/SS-B (extractable nuclear antigens) detected by enzyme-linked immunosorbent assay for the diagnosis of systemic lupus erythematosus. Arthritis Rheum. 1996. 39:1055–61.
16.Vos PA., Bast EJ., Derksen RH. Cost-effective detection of non-anti-double-stranded DNA antinuclear antibody specificities in daily clinical practice. Rheumatology. 2006. 45:629–35.
crossref

Fig. 1.
Results of fluorescent ANA (FANA), line immunoassay (LIA) and enzyme linked immunosorbent assay (ELISA) for anti-extractable nuclear antigen (ENA) antibodies in 437 patients. Numbers in parentheses represent the number of patients with systemic autoimmune diseases.
kjlm-28-353f1.tif
Table 1.
Clinical findings and test results of 29 patients with negative FANA and positive ENA antibodies by LIA
No. Case LIA (grade) ELISA Clinical diagnosis
1 SSA(3+) Negative Behcet's disease
2 SSA(3+) Negative Inflammatory arthralgia
3 SSA(3+) Positive Sjogren syndrome
4 SSA(3+) Positive Sjogren syndrome
5 SSA(3+) Positive Sjogren syndrome
6 SSA(3+) Positive Raynaud's syndrome
7 SSA(3+)/SSB(3+) Positive/Positive Sjogren syndrome
8 SSA(3+)/SSB(3+) Positive/Positive SLE
9 SSA(3+)/SSB(3+) Positive/Positive SLE
10 SSA(3+)/SSB(1+) Positive/Negative Raynaud's syndrome
11 SSA(3+)/SSB(1+) Positive/Negative Raynaud's syndrome
12 SSA(2+) Negative Rheumatoid arthritis
13 SSA(2+) Positive Rheumatoid arthritis
14 SSA(2+) Positive Arthritis
15 SSA(2+) Positive Adult onset Still's disease
16 SSA(1+) Negative Fibromyalgia
17 SSA(1+) Negative Fibromyalgia
18 SSA(1+) Negative Inflammatory arthralgia
19 SSA(1+) Negative Synovitis
20 SSA(1+) Positive Behcet's disease
21 SSA(1+) Negative Stomatitis
22 SSA(1+) Negative Stomatitis
23 SSA(1+) Negative Synovitis
24 SSB(1+) Negative Fibromyalgia
25 RNP(2+) Negative Osteoarthritis
26 RNP(1+) Negative Synovitis
27 Scl-70(1+) Negative Osteoarthritis
28 Jo-1(3+) Positive Polymyositis
29 Jo-1(1+) Negative Fibromyalgia

Abbreviations: FANA, fluorescent antinuclear antibodies; ENA, extractable nuclear antigen; LIA, line immunoassay; ELISA, enzyme linked immunosorbent assay; SLE, systemic lupus erythematosus.

Table 2.
Clinical findings and ELISA results of 22 patients with positive dsDNA antibody by LIA
No. Case FANA titer LIA (grade) ELISA Clinical diagnosis
1 Negative dsDNA(3+) Negative Reactive arthritis
2 Negative dsDNA(2+) Negative Behcet's disease
3 Negative dsDNA(2+) Negative Fibromyalgia
4 Negative dsDNA(2+) Negative Behcet's disease
5 Negative dsDNA(1+) Negative Osteoarthritis
6 Negative dsDNA(1+) Negative Osteoarthritis
7 Negative dsDNA(1+) Negative Gout
8 Homogenous 1:40 dsDNA(1+) Negative Inflammatory polyarthropathy
9 Homogenous 1:40 dsDNA(1+) Positive SLE
10 Speckled 1:80 dsDNA(2+) Positive SLE
11 Homogenous 1:80 dsDNA(1+) Negative Synovitis
12 Speckled 1:160 dsDNA(2+) Negative Rheumatoid arthritis
13 Speckled 1:160 dsDNA(1+) Negative Rheumatoid arthritis
14 Homogenous 1:160 dsDNA(1+) Negative Stomatitis
15 Homogenous 1:640 dsDNA(2+) Positive Overlap syndrome
16 Homogenous 1:640 dsDNA(1+) Positive SLE
17 Homogenous 1:1,280 dsDNA(2+) Positive SLE
18 Speckled 1:1,280 dsDNA(1+) Positive SLE
19 Speckled >1:1,280 dsDNA(2+) Positive SLE
20 Homogenous >1:1,280 dsDNA(2+) Positive Overlap syndrome
21 Homogenous >1:1,280 dsDNA(1+) Positive SLE
22 Homogenous >1:1,280 dsDNA(1+) Positive SLE

Abbreviations: See Table 1.

Table 3.
Agreement and discordances of LIA compared with ELISA in 53 patients with systemic autoimmune diseases
Group
(Cutoff of LIA=grade 1)		 No. of patients
SSA SSB RNP Sm Scl-70 Jo-1 dsDNA
ELISA+/LIA+ 23 14 10 4 3 1 12
ELISA+/LIA- 0 0 1 0 0 0 8
ELISA-/LIA+ 11 9 10 9 8 5 9
ELISA-/LIA- 19 30 32 40 42 47 24
Agreement (%) 79.2 83.0 79.2 83.0 84.9 90.6 67.9
PD (%) 36.7 23.1 23.8 18.4 16.0 9.6 27.3
ND (%) 0 0 9.1 0 0 0 40.0
Group
(Cutoff of LIA=grade 2)		 No. of patients
SSA SSB RNP Sm Scl-70 Jo-1 dsDNA
ELISA+/LIA+ 22 14 10 4 3 1 9
ELISA+/LIA- 1 0 1 0 0 0 10
ELISA-/LIA+ 5 4 5 2 2 1 2
ELISA-/LIA- 25 35 37 47 48 51 31
Agreement (%) 88.7 92.5 88.7 96.2 96.2 98.1 75.5
PD (%) 16.7 10.3 11.9 4.1 4.0 1.9 6.1
ND (%) 4.3 0 9.1 0 0 0 52.6

Agreement is defined as the concordance of the LIA with the ELISA. Positive discordance (PD) is calculated as the number of ELISA-/LIA+, divided by the total number of ELISA-. Negative discordance (ND) is calculated as the number of ELISA+/LIA-, divided by the total number ELISA+.

Abbreviations: ELISA, enzyme linked immunosorbent assay; LIA, line immunoassay; ND, negative discordance; PD, positive discordance.

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