Rapid screening and confirmatory methods for biochemical diagnosis of human prion disease

• Published in: Journal of virological methods 2011-08, Vol.175 (2), p.216-223
• Main Authors: Ugnon-Café, S., Dorey, A., Bilheude, J.M., Streichenberger, N., Viennet, G., Meyronet, D., Maues de Paula, A., Perret-Liaudet, A., Quadrio, I.
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Blotting, Western - methods; brain; Brain - pathology; Clinical Laboratory Techniques - methods; Creutzfeldt-Jakob Syndrome; Diagnosis; enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Fundamental and applied biological sciences. Psychology; Human; Humans; Mass Screening - methods; Microbiology; Palatine Tonsil - pathology; patients; Prion disease; Prion Diseases - diagnosis; PrPSc proteins; rapid methods; Retrospective Studies; screening; Sensitivity and Specificity; Spleen; Spleen - pathology; Techniques used in virology; Tonsil; tonsils; Transmissible spongiform encephalopathy; Virology; Western blotting
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2011.05.016

► We test commercial ELISA and Western blot originally designed for animal diagnosis. ► Both assays are suitable for human diagnosis in brain and lymphoid tissue. ► We obtain the expected molecular pattern of pathological prion protein. ► Commercial kits are robust assays and good alternative to time consuming methods. ► We propose a diagnostic strategy associating screening to confirmatory methods. Transmissible spongiform encephalopathies (TSEs) are characterised by accumulation of an abnormal isoform of prion protein (PrP sc), mainly in the brain but also in various peripheral tissues. Home-made assays consisting of non-standardised protocols are used currently for laboratory diagnosis of human TSE. The purpose of the present study was to test the ability of two commercial assays, TeSeE™ CJD ELISA and TeSeE™ Western blot, to detect PrPsc in cerebral and lymphoid tissues of TSE patients. Both tests detected a PrPsc-significant signal in the brains of 54 affected patients and not in 51 controls, yielding 100% specificity and 100% sensitivity. Furthermore, three post-mortem spleens and two pre-mortem tonsils from three patients with variant Creutzfeldt–Jakob disease (vCJD) were detected correctly. The expected PrPsc molecular patterns were found in TSE patient brain tissue and in the tonsils and spleens of the three vCJD patients. In conclusion, these rapid and robust in vitro tools were suitable for routine human TSE diagnosis and characterisation. CJD could also be diagnosed during the patient's lifetime by detection of PrPsc in the tonsil. A diagnostic strategy associating TeSeE™ CJD ELISA screening to biochemical confirmation by TeSeE™ Western blot is proposed.