DOI: 10.5858/arpa.2023-0274-oa ISSN: 1543-2165

C3d Immunohistochemical Staining on Paraffin-Embedded Tissue for Diagnosis of Pemphigus

Nanhui Wu, Yijie Cai, Fei Wu, Yulin Liang, Shuyi Liu, Pengfei Zhang, Yeqiang Liu
  • Medical Laboratory Technology
  • General Medicine
  • Pathology and Forensic Medicine


Pemphigus is an autoimmune blister disease that causes blisters on the skin and mucosal surfaces. Direct immunofluorescence (DIF) testing is critical for the clinical diagnosis of pemphigus. However, it is limited to fresh tissue specimens and fluorescence microscopy.


To assess the value of C3d immunohistochemistry (IHC) on paraffin-embedded skin tissue for the diagnosis of pemphigus by comparing C3d-IHC results to DIF and enzyme-linked immunosorbent assay testing in pemphigus and other blister-related skin diseases.


C3d-IHC assays were retrospectively performed on paraffin-embedded skin tissue sections from 115 patients (63 with pemphigus and 52 controls). Both the case group and the control group underwent the same protocol, and cases with C3d position in the peripheral spinous layer were considered as positive samples.


C3d-IHC and DIF testing had similar performance for pemphigus diagnosis, with a sensitivity of 71.0% (95% CI, 51.8%–85.1%) and 77.4% (95% CI, 58.5%–89.7%), specificity of 96.4% (95% CI, 79.8%–99.8%) and 100% (95% CI, 85.0%–100%), positive predictive value of 95.7% (95% CI, 76.0%–99.8%) and 100% (95% CI, 82.8%–100%), and a negative predictive value of 75.0% (95% CI, 57.5%–87.3%) and 80.0% (95% CI, 62.5%–90.9%), respectively.


Our study indicated that C3d-IHC results for paraffin-fixed tissues were not significantly different from DIF results for the diagnosis of pemphigus. The C3d-IHC assay has the potential for routine diagnosis of pemphigus, especially in the absence of fresh-frozen tissue.

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