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Immunohistochemical identification of HCV target antigen in paraffin-embedded liver tissue: reproducibility and
Gamal E Shiha1, Khaled R Zalata, Ahmed F Abdalla
1Department of Internal medicine, Mansoura University, Mansoura , 35516, Egypt. g_shiha@hotmail.com
Insights
Detecting hepatitis C virus (HCV) antigen in paraffin-embedded liver biopsies is now feasible. Immunohistochemical staining with TORDJI-22 MAb provides reproducible results in HCV-RNA positive patients.
Area of Science:
- Hepatology
- Virology
- Immunohistochemistry
Background:
- Immunohistochemical staining successfully detects hepatitis C virus (HCV) antigen in fresh frozen tissues.
- Limited and conflicting results exist for HCV antigen detection in paraffin-embedded tissues.
Purpose of the Study:
- To evaluate the identification of HCV antigen in paraffin-embedded liver biopsies.
- Utilize the anti-HCV monoclonal antibody (MAb) TORDJI-22 for antigen detection.
Main Methods:
- Applied immunohistochemical staining for HCV on 56 paraffin-embedded liver biopsy specimens.
- Included 46 HCV-RNA seropositive patients and 10 controls.
- Used TORDJI-22 MAb at 1:40 dilution with overnight incubation.
Main Results:
- Identified reproducible HCV antigen staining in 91% (42/46) of HCV-RNA seropositive cases.
- Observed a mainly coarse granular cytoplasmic staining pattern in hepatocytes, with occasional nuclear hue.
- Characterized three staining patterns (A, B, C); no similar pattern found in controls.
Conclusions:
- Immunohistochemical identification of HCV antigen is practical in paraffin-embedded liver biopsies with optimal techniques.
- The staining pattern is reproducible and primarily coarse granular cytoplasmic.
- No cross-reactivity with hepatitis B virus antigens was detected.
Background:
Immunohistochemical staining has been applied successfully to detect hepatitis C virus (HCV) antigen in fresh frozen tissue. In paraffin-embedded tissues, however, minimal trials with conflicting results have been reported.
Aims:
The present study is a trial to evaluate the identification of HCV antigen in paraffin-embedded liver biopsies using the anti-HCV monoclonal antibody (MAb) TORDJI-22.
Methods:
We applied immunohistochemical staining for HCV in 56 paraffin-embedded liver biopsy specimens, 46 from patients seropositive for HCV-RNA and 10 control liver biopsy specimens. The TORDJI-22 MAb was applied in dilution 1:40, with overnight incubation.
Results:
Reproducible staining patterns of HCV antigen in tissues were identified among the majority (42/46-91%) of HCV RNA seropositive cases. The staining pattern was cytoplasmic of hepatocytes, with occasional nuclear hue. It is mainly coarse granular with microvesicular pattern. Three staining patterns were identified: A, diffuse or membranous; B, patchy; and C, occasional paranuclear. None of the control samples showed a similar staining pattern.
Conclusion:
Immunohistochemical identification of HCV antigen is easy to apply in paraffin-embedded liver biopsy specimens when the optimal detection techniques are applied. The staining pattern is reproducible, being mainly coarse granular cytoplasmic. Cross reactivity with hepatitis B virus antigens was not detected.
