Milestone 1 Report – Project Implementation Plan
Pilot Study OneThe first pilot study will comprise eight PreservCyt specimens obtained from Victorian Cytology Service (VCS). Three specimens will be used and through diluting with PreservCyt solution, eight specimens will be produced. Two of these will be negatives and the remaining six will be positives of varying strengths (copy numbers) – see Figure 1. 20 laboratories will participate in this study. The focus of this method is establishing if the specimen result is indicative of high risk HPV.
A parallel study will be conducted with 5 laboratories using dry swabs. Eight specimens will be prepared by Sepehr Tabrizi. Cells will be inoculated onto flock swabs using human cell line C33A as it is readily available and HPV negative – see Figure 2. The focus of this method is sensitivity and specificity of the testing protocol.
Objective of Pilot Study OneThe main objective is to establish the best material and preparation method for specimens to issue to laboratories as a means of assessing laboratory performance.
Format:Approximately 20 laboratories will be enrolled to test PreservCyt specimens.
5mL of PreservCyt material will be sent to laboratories using the Digene Hybrid Capture method (most commonly in use) and 1mL of material will be sent to laboratories using alternative methods.
A parallel study will be conducted with 5 laboratories using dry swabs. There are a range of tests reporting HPV type and many more under development (especially in-house tests). WHO and CAP (College of American Pathologists) have used dry swabs and so it would be beneficial to compare the efficacy of the dry swab method with the PreservCyt method. The advantage of the dry swab is that the exact content of the specimen is known whereas, the exact content of the pooled PreservCyt solution is not known.
Therefore, it was decided that it would be beneficial to use the PreservCyt pools and then in addition, to send dry swabs to 5 laboratories. This would allow a more comprehensive assessment of the best mode for future QAP surveys and can be investigated further in Pilot Study 2.
Preparation for PreservCyt specimens (performed by Sepehr Tabrizi):Marion Saville from Victorian Cytology Service is located close to Sepehr Tabrizi at The Royal Women’s Hospital, Melbourne and is able to supply Sepehr with the required number of PreservCyt pools after they have been de-identified.
Development of an appropriate protocol will require working with the PreservCyts and testing by two different methods to ensure that dilution of the PreservCyt does not impact on any of the current testing protocols. Concerns are that once a single sample has been diluted, it may lose its cellularity which means that it is no longer a clinical sample. Mixing samples can create issues and so the feasibility of this method will require an extended “work-up” investigation and extensive pre-issue testing.
Preparation for dry swab specimens (performed by Sepehr Tabrizi)Top of pageCells will be inoculated onto a flock swab using the human cell line C33A as it is readily available and HPV negative. A panel of 8 swabs will be prepared with one prepared in duplicate for stability studies – the specimen that is around 50,000 copies/mL.
Participants will be asked to freeze this extra duplicate swab at -200C for two months and then test as a means for evaluating stability of specimens. [This will assess feasibility of sending all of the specimens in one dispatch in the future].
There is no existing protocol for Sepehr to follow and it is in effect a research project; therefore, it is difficult to define an exact time period of time for this work to be completed but it is anticipated that it would be completed by late March.
Paperwork Preparation (performed by the RCPA Serology QAP):Laboratories participating in the RCPA Serology QA program have already been notified of this impending Pilot Study and almost 20 have already expressed interest in participating. Flyers have now been forwarded to the RCPA Cytopathology QAP and they will distribute these to their participants.
In January each of the laboratories will be contacted and 20 chosen to be part of the first Pilot Study. Criteria will be the method that they use to detect HPV DNA, length of time that the laboratory has performed HPV testing and willingness to provide feedback and suggestions after each pilot study.
Method is important as it is helpful to have a wide range of methodologies to thoroughly check that the specimens are appropriate for all available methodologies.
Length of time that the laboratory has performed HPV testing is important as it is anticipated that laboratories that have been using this methodology for more than 12 months would have a more comprehensive understanding of the testing process than a laboratory that has recently introduced the methodology.
Willingness to provide feedback and suggestions after each pilot study is crucial to the successful development and implementation of this module.
The RCPA Serology QAP will enrol the laboratories in the Pilot Study and liaise with them regularly so that they are notified when specimens are ready for dispatch.
Questionnaire preparation will begin in January now that the type of specimen has been established. Website development will be based upon the format for the questionnaire. The RCPA Serology QAP website is well established and current data entry screens can be used as a template so that modifications can be made rather than starting from a blank screen. It is expected that these aspects of the preparation will be completed in February. Requirements for website development are: location for HPV questionnaires and reports; facility for data entry by participants; interactive graphical and statistical analysis on website and development of extra administrative functions relevant to HPV for such activities as the download of participant survey results.
PostScript to Milestone 1 ReportIt was anticipated that PreservCyt specimens would be obtained exclusively from Victorian Cytology Service (VCS). Due to a lack of sufficient high positive specimens, Symbion Laverty Pathology, North Ryde NSW provided numerous high positive specimen pools that were used in both Pilot Studies One and Two.Top of page
The questionnaire to ascertain method of testing, length of time that HPV testing has been performed HPV by each laboratory and the willingness of each participating laboratory to provide feedback and suggestions after each pilot study was prepared and distributed in February 2008 – see below.
Questionnaire issued to laboratories - 4th February, 2008 - Participant SurveyDownload participant survey (PDF 25 KB)
Summary of Results from Participant SurveyGeographical Distribution of Participants
- Australia x18 (7 NSW, 3 QLD, 3 VIC, 2 SA, 1 WA, 1 TAS)
- New Zealand x2
17 participants were existing SQAP participants, 3 were part of the Cytopathology program and showed interest following notification of a new HPV QA program
Distribution of Specimens
All 20 participants received 8 PreservCyt specimens
10 of these participants also received 8 Dry Swab specimens
- Australia x9 (4 NSW, 1 QLD, 2 VIC, 1 WA, 1 TAS)
- New Zealand x1
Purpose of HPV Testing
Period of Performing HPV Testing
Quantity of HPV Tests per month
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