Challenges in the Testing of Non-Heart-Beating Cadavers for Viral Markers: Implications for the Safety of Tissue Donors
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Ó Springer 2005
Challenges in the testing of non-heart-beating cadavers for viral markers: implications for the safety of tissue donors David Padley1, Morag Ferguson1,*, Ruth M Warwick2, Christopher Womack3, Sebastian B Lucas4 and John Saldanha1,5 1
Division of Virology, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Herts, EN6 3QG UK; 2Colindale Blood Centre, Colindale Avenue, London, NW9 5BG; 3Department of Cellular Pathology, Peterborough District Hospital, Thorpe Road, Peterborough, PE3 6DA; 4Department of Histopathology, St. Thomas’ Hospital, London SE1 7EH, UK; 5Current address: Roche Molecular Systems Inc., Pleasanton, California, USA; *Author for correspondence (e-mail: [email protected]; phone: +44-1707-641314; fax: +44-1707-646730) Received 29 October 2004; accepted in revised form 12 April 2005
Key words: Blood borne infection, Cadaver, HBV, HCV, HIV, NAT, Tissue bank, Viral marker
Abstract Natural changes that occur in blood and tissue after death may result in false positive results in antigen and antibody detection tests performed to identify markers of viral infection in potential tissue donors. Such tissue, which might otherwise be acceptable for therapeutic purposes, would not meet current standards for safe tissue banking. This is especially important in the context of insufficiency in the tissue supply. In this study, a series of blood samples collected during routine post-mortem examination was assayed using a range of commercially available kits for the detection of HBsAg, anti-HCV and anti-HIV 1 + 2 antibody/ antigen. Results of tests on 104 samples collected from 97 individuals indicate that some kits result in a higher number of initial reactive samples than others. Approximately 40% of samples were reactive in one or more HBsAg assay, less than 10% in at least one anti-HIV kit and only 1 sample at low level on an antiHCV kit. Liver or lymph node samples from individuals whose serum sample gave reactive results in antigen/antibody assays were tested for viral nucleic acid in the corresponding nucleic acid amplification test. Only one individual’s sample was confirmed to test positive for HBsAg in a confirmatory neutralisation test and by nucleic acid amplification technology, and a second individual whose serum was scored reactive for anti-HCV, but negative for HBsAg, had a liver sample which was HBV DNA positive and HCV RNA negative. The results of the study indicate that antibody/antigen assays are not as specific as NAT using state of the art DNA extraction techniques. Both types of assay complement each other and used together will help assure the safety of tissues for transplantation. Abbreviations: AIDS – Acquired immuno-deficiency syndrome; GZ – Grey zone; HBsAg – Hepatitis B surface antigen; HBV – Hepatitis B virus; HCV – Hepatitis C virus; HIV – Human immunodeficiency virus; NAT – Nucleic acid amplification techniques; RNA – Ribonucleic acid; RT-PCR – Reverse transcriptase polymerase chain reaction; DNA – Deoxy-ribonucleic acid
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