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Title: Development and application of nucleic acid amplification technology (NAT) for the detection of viruses in donated blood
Author: Grant, Paul Robert
ISNI:       0000 0001 3511 5869
Awarding Body: University of London
Current Institution: University College London (University of London)
Date of Award: 2003
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Background and Objectives: To reduce the potential for transmission of hepatitis C virus (HCV) from an RNA positive, anti-HCV negative blood donation, the National Blood Service (NBS) decided to introduce nucleic acid amplification technology (NAT) testing of blood donations for HCV in England and Wales. The objective of this thesis was to develop an automated assay using commercial components for the detection of HCV RNA in blood donations for transfusion, and to validate sample handling and storage procedures. Studies included in the thesis: The stability of HCV in the plasma sample was investigated to define the appropriate sample handling and storage conditions needed to preserve HCV RNA in the blood sample before analysis by NAT. HCV RNA was found to be more stable than previously thought, with HCV levels remaining stable for 120 hours in un-separated whole blood at refrigerated or room temperature. The compatibility of the Qiagen and Roche reagents were tested using manual assays and automated systems. This work has shown that it is possible to completely automate HCV NAT screening. Alternative techniques for virus detection such as TMA and HCV core antigen were also investigated and PCR was found superior to antigen and equivalent to TMA. The window period of the HCV infection was further defined in a study of window phase HCV samples from US plasma donors. Outcome: The combination of methods developed and described in this thesis were successfully introduced into routine use within the NBS from 1999 and are now used for release of blood components with a shelf life of greater than 24 hours in England and Wales. From 1999 to the end of 2002 approximately nine million donations were tested in England and Wales of which seven were HCV infected window phase units, which would have otherwise been transfused. In addition 394 NAT positive samples were found which were also detected by anti-HCV screening tests.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available