Anti-SS-A/Ro antibody determination by indirect immunofluorescence and comparison of different methods of anti-nuclear antibody screening

Noriyo Tanaka, Yoshinao Muro, Kazumitsu Sugiura, Yasushi Tomita

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Abstract

We evaluated the utility of HEp-2 cells transfected with the 60 kDa SS-A/Ro as a substrate for indirect immunofluorescence (IIF-HEp-2000) to compare several methods for screening Japanese serum samples for anti-SS-A/Ro antibodies. Serum samples from 243 Japanese patients were analyzed by IIF for anti-nuclear antibodies (ANAs), using HEp-2 cells (IIF-HEp-2), and for anti-SS-A/Ro 60 kDa antibodies, using IIF-HEp-2000 and enzyme-linked immunosorbent assay (ELISA). We performed double immunodiffusion and immunoprecipitation experiments, using the products of in vitro transcription and translation, to analyze sera for which there were discrepancies in the results of the IIF-HEp-2000 assay and ELISA. A total of 93 of the 243 serum samples showed findings positive for anti-SS-A/Ro antibodies. Notably, eight of the 93 sera gave positive findings for anti-SS-A/Ro antibodies by IIF-HEp-2000 but ANA-negative by IIF-HEp-2 analysis. Seven sera possessing anti-SS-A/Ro antibodies gave false negative results by IIF-HEp-2000; however, those samples were all ANA positive. ELISA for anti-SS-A/Ro antibodies showed that five and two samples gave false positive and negative results, respectively. Analysis by IIF-HEp-2000 was useful for primary screening of patients for ANAs, especially for anti-SS-A/Ro antibodies; the test could detect anti-SS-A/ Ro antibodies not identified on standard substrates in samples obtained from the Japanese population, as reported for the Caucasian population.

Original languageEnglish
Pages (from-to)585-592
Number of pages8
JournalModern Rheumatology
Volume18
Issue number6
DOIs
Publication statusPublished - 17-07-2008

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Indirect Fluorescent Antibody Technique
Anti-Idiotypic Antibodies
Serum
Enzyme-Linked Immunosorbent Assay
SS-A antibodies
Immunodiffusion
Immunoprecipitation
Population

All Science Journal Classification (ASJC) codes

  • Rheumatology

Cite this

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title = "Anti-SS-A/Ro antibody determination by indirect immunofluorescence and comparison of different methods of anti-nuclear antibody screening",
abstract = "We evaluated the utility of HEp-2 cells transfected with the 60 kDa SS-A/Ro as a substrate for indirect immunofluorescence (IIF-HEp-2000) to compare several methods for screening Japanese serum samples for anti-SS-A/Ro antibodies. Serum samples from 243 Japanese patients were analyzed by IIF for anti-nuclear antibodies (ANAs), using HEp-2 cells (IIF-HEp-2), and for anti-SS-A/Ro 60 kDa antibodies, using IIF-HEp-2000 and enzyme-linked immunosorbent assay (ELISA). We performed double immunodiffusion and immunoprecipitation experiments, using the products of in vitro transcription and translation, to analyze sera for which there were discrepancies in the results of the IIF-HEp-2000 assay and ELISA. A total of 93 of the 243 serum samples showed findings positive for anti-SS-A/Ro antibodies. Notably, eight of the 93 sera gave positive findings for anti-SS-A/Ro antibodies by IIF-HEp-2000 but ANA-negative by IIF-HEp-2 analysis. Seven sera possessing anti-SS-A/Ro antibodies gave false negative results by IIF-HEp-2000; however, those samples were all ANA positive. ELISA for anti-SS-A/Ro antibodies showed that five and two samples gave false positive and negative results, respectively. Analysis by IIF-HEp-2000 was useful for primary screening of patients for ANAs, especially for anti-SS-A/Ro antibodies; the test could detect anti-SS-A/ Ro antibodies not identified on standard substrates in samples obtained from the Japanese population, as reported for the Caucasian population.",
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Anti-SS-A/Ro antibody determination by indirect immunofluorescence and comparison of different methods of anti-nuclear antibody screening. / Tanaka, Noriyo; Muro, Yoshinao; Sugiura, Kazumitsu; Tomita, Yasushi.

In: Modern Rheumatology, Vol. 18, No. 6, 17.07.2008, p. 585-592.

Research output: Contribution to journalArticle

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T1 - Anti-SS-A/Ro antibody determination by indirect immunofluorescence and comparison of different methods of anti-nuclear antibody screening

AU - Tanaka, Noriyo

AU - Muro, Yoshinao

AU - Sugiura, Kazumitsu

AU - Tomita, Yasushi

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