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Peptide-based ELISAs are not sensitive and specific enough to detect muscarinic receptor type 3 autoantibodies in serum from patients with Sjögren's syndrome
  1. N Roescher1,2,
  2. A Kingman1,
  3. Y Shirota1,
  4. J A Chiorini1,
  5. G G Illei1
  1. 1National Institutes of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA
  2. 2Academic Medical Center, Amsterdam, The Netherlands
  1. Correspondence to Nienke Roescher, NIH/NIDCR/MPTB, 10 Center Drive, Building 10, Room 1A21, Bethesda, MD 20892-1190, USA; roeschern{at}mail.nih.gov

Abstract

Objectives The detection of autoantibodies to the muscarinic receptor type 3 (M3R) in the serum of patients with Sjögrens syndrome (SS) by ELISA is controversial. A study was undertaken to test whether modification of M3R peptides could enhance the antigenicity and increase the detection of specific antibodies using an ELISA.

Methods A series of controlled ELISAs was performed with serum from 71 patients with SS and 37 healthy volunteers (HV) on linear, citrullinated and/or cyclised and multi-antigenic peptides (MAP) of the three extracellular M3R loops to detect specific binding.

Results Significant differences (p<0.05) in optical density (OD) between serum from patients and HV were detected for a cyclised loop 1-derived peptide and the negative control peptide. Furthermore, there were no statistically significant differences between the frequency of positive patients (defined as OD >2SDs above the mean of the HV) and HV on any of the peptides tested.

Conclusions Binding of serum from patients with SS to M3R-derived peptides does not differ from binding to a control peptide in an ELISA and no significant binding to M3R-derived peptides was found in the serum from individual patients compared with HV. These data suggest that peptide-based ELISAs are not sufficiently sensitive and/or specific to detect anti-MR3 autoantibodies.

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Footnotes

  • Funding This research was supported by the intramural research program of the NIH, NIDCR.

  • Competing interests None.

  • Ethics approval All subjects signed an informed consent and the study was approved by the Institutional Review Board (IRB) of the National Institutes of Dental and Craniofacial Research (NIDCR).

  • Provenance and peer review Not commissioned; externally peer reviewed.

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