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Korean J Lab Med. 2010 Dec;30(6):675-684. Korean.
Published online December 02, 2010.  https://doi.org/10.3343/kjlm.2010.30.6.675
Copyright © 2010 The Korean Society for Laboratory Medicine
Evaluation of Anti-dsDNA Antibody Tests: Crithidia luciliae Immunofluorescence Test, Immunoblot, Enzyme-linked Immunosorbent Assay, Chemiluminescence Immunoassay
Jin-young Yang, M.D., Eun-Jee Oh, M.D., Yonggoo Kim, M.D. and Yeon-Joon Park, M.D.
Department of Laboratory Medicine, The Catholic University of Korea School of Medicine, Seoul, Korea.

Corresponding author: Eun-Jee Oh, M.D. Department of Laboratory Medicine, Seoul St. Mary's Hospital, 505 Banpo-dong, Seocho-gu, Seoul 137-701, Korea. Tel: +82-2-2258-1641, Fax: +82-2-2258-1719, Email: ejoh@catholic.ac.kr
Received May 31, 2010; Revised August 18, 2010; Accepted October 09, 2010.

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, 3 ELISA 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.

Keywords: Anti-dsDNA; Crithidia luciliae immunofluorescence test; ELISA; Chemiluminescence immunoassay; Immunoblot assay

Figures


 Fig. 1
Anti-dsDNA levels measured by ELISA I (A), II (B), III (C), and CLIA (D) in 74 SLE patients, 50 patients with other systemic rheumatic diseases (ORD) and 18 patients with other diseases (Others) (Cut-off I provided by manufactures, cut-off II determined by ROC curve, and cut-off III set at 95% specificity are indicated.).

Abbreviations: SLE, systemic lupus erythematosus; ORD, other systemic rheumatic diseases; IB, immunoblot assay; ELISA, enzyme-linked immunosorbent assay; CLIA, chemiluminescence immunoassay.
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Fig. 2
ROC curve of the 3 ELISA assay kits and one CLIA.

Abbreviations: ELISA, enzyme-linked immunosorbent assay; CLIA, chemiluminescence immunoassay.
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Fig. 3
The correlation among three commercial ELISA and one CLIA kits for anti-dsDNA detection in 142 serum samples.

Abbreviations: ELISA, enzyme-linked immunosorbent assay; CLIA, chemiluminescence immunoassay.
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Tables


 Table 1
Characteristics of assay kits used for the detection of anti-dsDNA
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Table 2
Analytical performance of various anti-dsDNA antibody assays based on the manufacturers' cut-off values
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Table 3
Analytical performance of various anti-dsDNA antibody assays based on the cut-off values determined by ROC curve analysis
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Table 4
Analytical performance of various anti-dsDNA antibody assays based on the cut-off values set at 95% specificity by ROC curve analysis
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Table 5
Concordance rates (%) of anti-dsDNA detection results among different assay kits based on various cut-off values
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Notes

This work was supported by a grant of the Korea Healthcare technology R & D project, Ministry for Health, Welfare & Family Affairs, Republic of Korea (A092258).

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