Evaluation of Anti-dsDNA Antibody Tests: Crithidia luciliae Immunofluorescence Test, Immunoblot, Enzyme-linked Immunosorbent Assay, Chemiluminescence Immunoassay

다국어 초록 (Multilingual Abstract)

Background : Anti-double stranded DNA antibody (anti-dsDNA) test is useful for the diagnosis and monitoring of systemic lupus erythematosus (SLE). Although several methods are available, none of them is completely satisfactory and differences among them have been reported. We evaluated the diagnostic performance of 6 commercial kits for anti-dsDNA detection.
Methods : A total of 142 sera (SLE [N=74], other systemic rheumatic diseases [N=50], other diseases [N=18]) were tested by 6 different assay kits using different antigenic sources of DNA: Crithidia luciliae immunofluorescence test (CLIFT), salmon testes (immunoblot, IB), human (ELISA I), salmon testes with nucleosome linker (ELISA II), plasmid (ELISA III), and synthetic oligonucleotides (chemiluminescence immunoassay, CLIA).
Results : With manufacturers’ cut-off values, 6 test kits showed sensitivities of 55.4-91.9%. ELISA I had a greater sensitivity than the other five assays (P<0.001). The specificities of ELISA II, ELISA III, CLIA, and CLIFT were higher than those of ELISA I and IB (P<0.05). In ROC curve analysis, 3ELISA kits and CLIA showed AUC values of 0.845-0.893, and revealed no significant differences among them (P>0.05). With cut-off values set at 95% of specificity, ELISA II had a higher sensitivity than ELISA III (63.5% vs. 41.9%, P<0.05). IB had poor concordance rates with other assays (42.0-65.0%). Pearson correlation coefficients among 4 quantitative assays were 0.667-0.798.
Conclusions : Six different assays showed various performances depending on the methods and cut-off values used. Except IB, the other five assays can be used for the detection of anti-dsDNA. (Korean J Lab Med 2010;30:675-84)

참고문헌 (Reference)

1 김지명, "전신 자가면역질환에서 Line Immunoassay를 이용한 항 ENA항체 및 항 dsDNA항체의 검출" 대한진단검사의학회 28 (28): 353-361, 2008

2 Sinico RA, "The use of laboratory tests in diagnosis and monitoring of systemic lupus erythematosus" 15 (15): S20-S27, 2002

3 Feltkamp TE, "The first international standard for antibodies to double stranded DNA" 47 : 740-746, 1988

4 Tan EM, "The 1982 revised criteria for the classification of systemic lupus erythematosus" 25 : 1271-1277, 1982

5 López-Longo FJ, "Simultaneous identification of various antinuclear antibodies using an automated multiparameter line immunoassay system" 12 : 623-629, 2003

6 Wasmuth JC, "ROC analysis comparison of three assays for the detection of antibodies against double-stranded DNA in serum for the diagnosis of systemic lupus erythematosus" 50 : 2169-2171, 2004

7 Reveille JD, "Predictive value of autoantibodies for activity of systemic lupus erythematosus" 13 : 290-297, 2004

8 Smeenk RJ, "Methodological update detection of antibodies to ds-DNA: current insights into its relevance" 20 : 294-300, 2002

9 Wong KH, "Measurement of anti-dsDNA: a comparative study of two ELISA and the Crithidia assay" 30 : 57-61, 1998

10 Nossent JC, "Low avidity antibodies to dsDNA as a diagnostic tool" 48 : 748-752, 1989

11 Sinico RA, "Interdisciplinary forum on autoimmune disease research:guidelines" 19 : 212-224, 2002

12 Tozzoli R, "Guidelines for the laboratory use of autoantibody tests in the diagnosis and monitoring of autoimmune rheumatic diseases" 117 : 316-324, 2002

13 Deng JS, "Gilliam JN. The binding of antihistone antibodies to Crithidia luciliae kinetoplasts is growth cycledependent" 28 : 163-168, 1985

14 Villalta D, "Evaluation of a new automated enzyme fluoroimmunoassay using recombinant plasmid dsDNA for the detection of anti-dsDNA antibodies in SLE" 16 : 227-232, 2002

15 Bencivelli W, "Disease activity in systemic lupus erythematosus: report of the consensus study group of the European workshop for rheumatology research. III. Development of a computerised clinical chart and its application to the comparison of different indices of disease activity. The European Consensus Study Group for Disease Activity in SLE" 10 : 549-554, 1992

16 Antico A, "Diagnosing systemic lupus erythematosus: new-generation immunoassays for measurement of anti-dsDNA antibodies are an effective alternative to the Farr technique and the Crithidia luciliae immunofluorescence test" 19 : 906-912, 2010

17 Rouquette AM, "Detection of antibodies to dsDNA:an overview of laboratory assays" 15 : 403-407, 2006

18 Janyapoon K, "Detection of anti-dsDNA by ELISA using different sources of antigens" 37 : 63-68, 2005

19 Horák P, "Clinical utility of selected disease activity markers in patients with systemic lupus erythematosus" 20 : 337-344, 2001

20 Kim KH, "Clinical significance of ELISA positive and immunofluorescence negative anti-dsDNA antibody" 380 : 182-185, 2007

21 Isenberg D, "Clinical laboratory assays for measuring anti-dsDNA antibodies. Where are we now?" 11 : 797-800, 2002

22 Hylkema MN, "Clinical evaluation of a modified ELISA, using photobiotinylated DNA, for the detection of anti-DNA antibodies" 170 : 93-102, 1994

23 Haugbro K, "AntidsDNA antibodies and disease classification in antinuclear antibody positive patients: the role of analytical diversity" 63 : 386-394, 2004

24 Blatt NB, "Anti-DNA autoantibodies and systemic lupus erythematosus" 83 : 125-139, 1999

25 Kavanaugh AF, "American College of Rheumatology Ad Hoc Committee on Immunologic Testing Guidelines. Guidelines for immunologic laboratory testing in the rheumatic diseases: anti-DNA antibody tests" 47 : 546-555, 2002

26 Radice A, "A new oligonucleotide-based ELISA for the detection of anti-double-stranded DNA antibodies" 39 : 113-119, 2006

27 Emlen W, "A new ELISA for the detection of double-stranded DNA antibodies" 132 : 91-101, 1990

28 Tan EM, "A critical evaluation of enzyme immunoassays for detection of antinuclear autoantibodies of defined specificities. I. Precision, sensitivity, and specificity" 42 : 455-464, 1999

29 Fritzler MJ, "A critical evaluation of enzyme immunoassay kits for detection of antinuclear autoantibodies of defined specificities. III. Comparative performance characteristics of academic and manufacturers’ laboratories" 30 : 2374-2381, 2003