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Published on: April 8, 2016
A non-biotin polymerized horseradish-peroxidase method for the immunohistochemical diagnosis of canine distemper
C T Liang1, L L Chueh, V F Pang
1National Laboratory Animal Center, National Applied Research Laboratories, Nan-Kang, Taipei 115.
Abstract:
This report describes a modified non-biotin polymerized horseradish peroxidase (HRP) immunohistochemical method for the diagnosis of canine distemper virus (CDV) infection from formalin-fixed, paraffin wax-embedded tissues. This method confirmed infection in seven of eight (87.5%) suspected cases. Labelled CDV antigen was observed in the following sites: cerebrum, cerebellum, meninges, glial cells, neurons, vascular endothelium, periventricular areas and pericytes, and choroid plexus; grey and white matter and central canal of the spinal cord; renal pelvis and tubular epithelium, and urinary bladder epithelium; macrophages and lymphocytes in splenic white pulp and lymph nodes; skin epidermis; bronchiolar epithelium and alveolar macrophages; hepatic Kupffer cells, and gastric and intestinal mucosal epithelium; stratified squamous epithelium of the tongue and oesophagus. With the non-biotin HRP detection system, pretreatment by autoclaving followed by microwave heating gave better labelling results than did microwave pretreatment alone. No obvious difference was noted between the labelling results produced by the non-biotin HRP detection system and the Super Sensitive Link-Label IHC detection system.
Insights
A new immunohistochemical method effectively diagnoses canine distemper virus (CDV) infection in tissues. This non-biotin polymerized horseradish peroxidase (HRP) technique showed high sensitivity in suspected cases.
Area of Science:
- Veterinary Pathology
- Immunohistochemistry
- Virology
Background:
- Canine distemper virus (CDV) poses a significant threat to canine health.
- Accurate diagnosis of CDV infection is crucial for effective disease management.
- Traditional diagnostic methods may have limitations in sensitivity or specificity.
Purpose of the Study:
- To develop and validate a modified non-biotin polymerized horseradish peroxidase (HRP) immunohistochemical method for CDV diagnosis.
- To assess the diagnostic performance of the new method in formalin-fixed, paraffin-embedded canine tissues.
- To optimize pretreatment protocols for enhanced antigen detection.
Main Methods:
- Utilized a modified non-biotin polymerized HRP immunohistochemical technique.
- Tested the method on formalin-fixed, paraffin wax-embedded tissues from suspected CDV cases.
- Compared different pretreatment methods (microwave alone vs. autoclaving followed by microwave heating).
- Evaluated labeling results against the Super Sensitive Link-Label IHC detection system.
Main Results:
- The method confirmed CDV infection in 87.5% (7/8) of suspected cases.
- CDV antigen was detected in multiple tissues, including brain, spinal cord, kidneys, lymph nodes, spleen, skin, lungs, liver, gastrointestinal tract, tongue, and esophagus.
- Autoclaving followed by microwave heating yielded superior labeling compared to microwave pretreatment alone.
- No significant difference in labeling was observed between the non-biotin HRP system and the Super Sensitive Link-Label IHC system.
Conclusions:
- The modified non-biotin HRP immunohistochemical method is a sensitive and reliable tool for diagnosing canine distemper virus infection.
- The optimized pretreatment protocol enhances antigen detection, improving diagnostic accuracy.
- This method provides comprehensive information on CDV distribution within affected tissues.
