Department of Veterinary Medicine

Research in Veterinary Medicine

Research description

We are making progress towards the development of simple diagnostics tests for detecting transmissible spongiform encephalopathies in sheep, cattle and humans. In particular, we have developed a unique Mab (YWH1) that is operationally specific for the disease-associated form of prion protein (PrP) in tissues. Accordingly, a very simple one step extraction assay has been developed for ovine brain, spinal cord, palatine tonsil and other lymphoid tissue that clearly differentiates between normal and histology-confirmed scrapie-positive animals without the need for proteinase K as a discriminating reagent. Currently, we are collaborating with Enfer Scientific to commercialise this procedure and to participate in a full evaluation conducted by the European Commission. In addition, we have produced a second reagent (YWH2) using phage display technology which recognises the equivalent human PrP sequence. In collaboration with the Health Protection Agency, we are investigating the suitability of the YWH2 protocol to service the human tonsil survey. We are also applying the methods to the detection of disease-associated PrP in more readily available tissues, blood fractions and urine within a pre-clinical context to determine the earliest time when infection can be identified and tracking pathogenesis through to its terminal stage.

Main collaborators

• John Coward, RevelationBio Ltd.
  426-428 Holdenhurst Road, Bournemouth Dorset BH8 9AA
• Drs. John Clarke and Amanda O'Brien, Enfer Scientific
  Unit T, M7 Business Park, Newhall, Naas, Co. Kildare, Ireland
• Dr. Reza Dabaghian, Health Protection Agency
  Centre for Infections 61 Colindale Avenue London NW9 5EQ
• Drs. Jim Hope and Maurice Sauer, Veterinary Laboratories Agency
  New Haw Addlestone Surrey KT15 3NB
• Dr. Balbir Raj, Unipath Limited, Priory Business Park, Bedford MK44 3UP
• Professor Man-Sun Sy, Professor of Pathology and Neurosciences
  School of Medicine, Case Western Reserve University
  10900 Euclid Ave, Cleveland OH 44120 USA
• Duncan Talbot, Unilever Corporate Research
  Colworth House, Sharnbrook, Bedfordshire MK44 1LQ
• Dr. Fortune Kohen, Department of Biological Regulation
  Weizmann Institute of Science, Rehovot, Israel

Key publications since 2001

• Barnard G, Hopkins L, Moorthie S, Seilly D, Tonks P, Dabaghian R, Clewley JP, Coward J, McConnell I (2007) Direct detection of disease associated prions in brain and lymphoid tissue using antibodies recognizing the extreme N-terminus of PrPC. Prion, 1:121-127.
• Talbot DCS, Ogborne RM, Dadd T, Adlercreutz H, Barnard G, Bugel S, Kohen F, Marlin S, Piron J, Cassidy A, Powell J (2007) Monoclonal Antibody-Based Time-Resolved Fluorescence Immunoassays for Daidzein, Genistein, and Equol in Blood and Urine: Application to the Isoheart Intervention Study. Clinical Chemistry, 53:748-756.
• Dabaghian, R, Barnard G, McConnell I, Clewley JP (2006) An immunoassay for the pathological form of the prion protein based on denaturation and time resolved fluorometry. Journal of Virological Methods 132:85-91.
• Brabin L, Roberts SA, Fairbrother E, Mandal D, Higgins SP, Chandiok S, Wood P, Barnard G, Kitchener HC (2005) Factors affecting vaginal pH levels among female adolescents attending genitourinary medicine clinics. Sexually Transmitted Infections 81:483-487.
• Pan T, Chang B, Wong P, Li C, Li R, Kang S-C, Robinson JD, Thompsett AR, Tein P, Yin S, Barnard G, McConnell I, Brown DR, Wisniewski T, Sy M-S (2005) An Aggregation-Specific Enzyme-Linked Immunosorbent Assay: Detection of Conformational Differences between Recombinant PrP Protein Dimers and PrPSc Aggregates. Journal of Virology 79: 12355-12364.
• Brabin L, Fairbrother E, Mandal D, Roberts SA, Higgins SP, Chandiok S, Wood P, Barnard G, Kitchener HC (2005) Biological and hormonal markers of chlamydia, human papillomavirus and bacterial vaginosis among adolescents attending genitourinary medicine clinics. Sexually Transmitted Infections 81:128-132.
• Minor P, Newham J, Jones N, Bergeron C, Gregori L, Asher D, van Engelenburg F, Stroebel T, Vey M, Barnard G, Head M and the WHO Working Group on International Reference Materials for the Diagnosis and Study of Transmissible Spongiform Encephalopathies (TSEs) (2004) Standards for the assay of Creutzfeldt-Jakob disease specimens. Journal of General Virology 85:1777-1784.
• Kang S-C, Li R, Wang C, Pan T, Liu T, Rubenstein R, Barnard G, Wong B-S, Sy M-S (2003) Guanidine hydrochloride extraction and detection of prion proteins in mouse and hamster by ELISA. Journal of Pathology 199: 534-541.
• Barnard G, Sy M-S (2003) The Diagnosis of Transmissible Spongiform Encephalopathies using Differential Extraction and DELFIA. In: Nunnally B, Krull I (eds.) Prions and Mad Cow Disease. New York: Marcel Dekker, pp. 277-315.