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Request and Response for details of UK 2010 XMRV blood donor study, from the UK Healt

Discussion in 'XMRV Research and Replication Studies' started by Wildcat, Mar 17, 2011.

  1. Wildcat

    Wildcat Senior Member

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    Request and Response for details of UK 2010 XMRV blood donor study, from the UK Health Protection Agency under the UK Freedom of Information Act.



    'In response to a question by Caroline Lucas in the Commons (18th October 2010), Under Secretary of State for Health Anne Milton stated:

    An expert subgroup of National Expert Panel for New and Emerging Infections (NEPNEI) met in May 2010, to consider all available evidence about XMRV and conduct a risk assessment. and The NHS Blood and Transplant and Health Protection Agency study group concur with the views expressed both by NEPNEI and SaBTO but also recognise the need for further research on the prevalence of XMRV in the United Kingdom. In a recent unpublished pilot study conducted by the group a series of 540 randomly selected English blood donors were screened for XMRV and none were found to be infected.
    (Hansard 18 Oct 2010 : Column 566W)



    Please send me all information relating to the XMRV screening pilot study on 540 English blood donors referred to by Anne Milton, including, but not restricted to:

    1. Study design and protocol.

    2. Who the Principal Investigators are.

    3. All collaborating institutions.

    4. Specific details of the specific test methods used for detection of XMRV in blood donors (eg specifics of the PCR, culture tests, serology etc).

    5. Where the blood drawing was carried out and by whom.

    6. Where the blood testing (PCR and other associated techniques) was carried out and by whom.

    7. Where the analysis of the results was carried out and by whom.

    8. If this study is intended for publication, and if so when.'





    ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~




    Response from the UK Health Protection Agency under the UK Freedom of Information Act, 7th March 2011:


    The following are responses to the questions asked above. These responses relate to the screening pilot study of 540 English blood donors as requested.


    1. The study design was to determine the prevalence of the XMRV provirus in blood donors in England using the published TaqMan protocol described by McCormick et al. [1] and the protocol was as follows:


    Random anonymous whole blood samples were obtained from Donation Testing at the National Health Service Blood and Transplant (NHSBT) centre at Colindale, London NW9.


    Initially for the first 78 samples, pellets of buffy were prepared and stored at 30C prior to extraction. For the next 129 samples buffy coat was prepared and reconstituted with PBS to the original blood volume and stored frozen at 30C prior to extraction. Finally 333 fresh whole blood samples were tested without fractionation.


    Nucleic acid was extracted from all samples on a Qiagen MDx Biorobot using the QIAamp One For All Kit (Catalogue number 965672) and the QIAamp One For All MDx cV70a or cV60a protocol using 273 l of sample and eluted in 80 l of Qiagen AVE buffer.


    10 l of the nucleic acid extract was analysed separately in three individual quantitative PCRs (Q-PCRs). One Q-PCR tested for XMRV, one Q-PCR tested for a co-extracted soil borne cereal mosaic virus (SBCMV) DNA control (internal control) and the other Q-PCR measured the input of human DNA (Pyruvate dehydrogenase (PDH) gene, cell input control). Samples invalid on either control were excluded from the analysis.


    If a sample signalled on initial XMRV testing it was re-extracted and re-tested. To be considered positive it had to re-signal on repeat XMRV Q-PCR testing.



    2. Professor Richard Tedder


    3. Health Protection Agency, NHS Blood and Transplant,


    4. Specific details of the specific test methods used for detection of XMRV in blood donors eg specifics of the PCR


    The quantitative PCR used to search for XMRV (and related Murine Leukaemia Virus) proviral sequences in the DNA from blood donors was a published TaqMan assay (McCormick, Brown et al. 2008) but performed using reagents as detailed in Table 1, as these proved optimal in our experiments. PCRs were performed in a 25 l volume on 10 l of sample with 400nM concentrations of primers and 200nM probes.


    Table 1
    PCR Primers and probes Reagents Conditions
    XMRV Taq Man XMRV Probe, F, R (McCormick, Brown et al. 2008)
    Qiagen Quanti Tect Probe kit 15 min at 95 C
    (15 secs 95 C, 1 min 60C) 60 cycles.



    5. In accordance with section 1 (1) (a) of the Freedom of Information Act I confirm that we do not hold this information as these were random anonymised donors.


    6. At Microbiology Services (formerly Centre for Infections) at the Health Protection Agency, Colindale, London. The work was performed by members of Professor Tedders team.


    7. As above



    8. The results of this study will be added to the results of on-going work. Once all of the work is complete we intend to publish our work. We anticipate this happening in the first half of this year.



    References

    McCormick, A. L., R. H. Brown, Jr., et al. (2008). "Quantification of reverse transcriptase in ALS and elimination of a novel retroviral candidate." Neurology 70(4): 278-283.




    I hope you have found this information useful, however, if you are dissatisfied with this response and would like a copy of the HPA complaints procedure then please contact Mr George Stafford, Complaints Manager at: Health Protection Agency, 61 Colindale Avenue, London NW9 5EQ.

    Please note that you have the right to an independent review by the Information Commissioners Office if a complaint cannot be resolved through the HPA complaints procedure. The Information Commissioners Office can be contacted by writing to Information Commissioners Office, Wycliffe House, Water Lane, Wilmslow, Cheshire SK9 5AF.

    Please contact me if you require any further information or assistance.

    Yours sincerely
    Freedom of Information Officer
    Health Protection Agency

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