USER MANUAL - Scotland's Health on the Web
Contents
1. 3 Review date 10 09 2016 Protocol for sectioning paraffin blocks at source laboratory It is the responsibility of the sending laboratory to ensure that the relevant block which contains the lesion of interest is selected 1 Ensure microtome and surrounding area are cleaned thoroughly recommended cleaner microsol wipes and that howie coat and gloves are worn throughout the procedure 2 Select the relevant FFPE block and face up to ensure even surface 3 Using a fresh blade take 10 um section and place into a 2 ml DNAse RNAse free screw cap tube eg sarstedt tube or eppendorf avoid using cryo vials or wider diameter tubes if possible 4 Using a new blade take and additional 10 um section and place into a second Screw cap sarstedt tube 5 Tubes containing the section should be labelled with Surname Forename CHI and DOB as instructed in section 5 2 If sending a section to SHPVRL for the first time use an additional third new blade to section a blank paraffin block containing no tissue and place in a separate tube This will be tested to exclude potential environmental contamination If a regular requestor you may be asked to send additional blank sections intermittently SHPVRL will contact you with details Jot 29 Author K Cuschieri Johannessen amp C Moore2. 59 66 and 68 The Abbott assay is based on PCR based amplification of target DNA in the sample 8 positive result would indicate that the assay has detected at least one of the types listed above a negative result would indicate that none of the types listed above are present in the sample or are present at levels below the detection limit of the assay The assay also contains an internal cellular control to check for specimen adequacy to minimise the possibility of false negatives 8 Jof 29 Author K Cuschieri Johannessen amp C Moore Doc no HPV 64 VERSION 4 Review date 10 09 2016 The results that will be available in SCCRS will be Result in SCCRS Definition HR HPV Positive Positive for 1 or more of the 14 high risk types listed above HR HPV Negative Negative for any of the high risk types listed above HPV Fail Issue with the sample sample processing or test meaning definitive result could not be generated These are the only definitive results that will map to letters Factors that could affect test performance or interpretation The Abbott assay requires LBC sample To allow for slight excess repeat and for volume loss to pipetting laboratories are required to provide a 3 ml prequot of LBC sample The term prequot refers to an aliquot that is removed from the original LBC vial before it is transferred to an automated slide processor At time of preparation of this SOP we do not recommend that samples which have
3. Cuschieri Johannessen amp C Moore Doc no HPV 64 VERSION 4 Review date 10 09 2016 ROO ES Information for Scottish Cytology Labs HPV Testing as a test of cure of treatment HPV testing for women who have been treated for cervical lesions as a test of cure rolled out across Scotland from April 3Oth 2012 HPV testing for test of cure occurs centrally at the Scottish HPV Reference Laboratory The following section outlines e Key information about the HPV test used e Guidance on sample prequot preparation and secure specimen transfer e Information on the HPV test used for test of cure and the associated results THE HPV TEST AND ASSOCIATED RESULTS At time of preparation of this document the test that is used for Test of Cure is the Abbott RealTime High Risk HPV Test This is a clinically validated test that has Certificate Europeanne CE marking and satisfies non inferiority criteria of Meijer et al 2009 with respect to diagnostic HPV assays It is also one of the assays approved by the English Cervical Screening Programme for triage and test of cure The Abbott assay tests for a group of 14 high risk HPV types in aggregate see detection range below it also provides additional limited typing of HPV 16 and 18 although type specific result will not be routinely entered in SCCRS and management is according to HR HPV positive or HR HPV negative status Detection range 16 18 31 33 35 39 45 51 52 56 58
4. as described above Jof 28 If the original LBC container is sent please ensure it is not required for any aspect of the cytopathology process at the sending laboratory SHPVRL is not responsible for returning original LBC samples to the sending laboratory If prequot is sent ensure container is labelled according to section 5 2 Furthermore original vials should be vortexed prior to prequot or manually inverted 6 times if no vortex available All prequots should made using sterile plasticware Order details for prequot tubes are in section 6 0 If sending paraffin sections it is essential to refer to Protocol for sectioning paraffin blocks at source laboratory in this user manual Paraffin sections of tissue in other fixatives eg Bouin s solution and fresh tissue are not generally acceptable for HPV testing for further information contact SHPVRL SHPVRL will send all original blocks back to parent laboratory If the original LBC container is sent please ensure it is not required for any aspect of the cytopathology process at the sending laboratory SHPVRL is not responsible for returning original LBC samples to the sending laboratory If prequot is sent for an individual clinical request ensure container is labelled according to section 5 2 All prequots should made using sterile plasticware Order details for prequot tubes are in section 6 0 pted after prior discussion with SHPVRL Author K
5. research collaborations In pursuit of the above staff at SHPVRL work closely with the HPV Research Group in the Queens Medical Research Group University of Edinburgh Division of Pathology which hosts the National HPV archive a biobank of over 20 000 samples set up to facilitate HPV research The archive welcomes applications for sample use from interested researchers for further information please contact hpvarchive ed ac uk SHPVRL was instrumental in the creation of the Scottish HPV Investigators Network SHINe and contributes to its continuing activities SHINe is a cross disciplinary forum designed to engender and deliver collaborative research on HPV For further details please see the SHINe Website www shine mvm ed ac uk of 29 Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 Review date 10 09 2016 amp 45 SELECTED PUBLICATIONS 1 Pollock KG Kavanagh K Potts A Love J Cuschieri K Cubie H Robertson C Cruickshank M Palmer TJ Nicoll S Donaghy M Reduction of low and high grade cervical abnormalities associated with high uptake of the HPV bivalent vaccine in Scotland Br J Cancer 2014 Sep doi 101038 bjc 2014 479 2 Ward C Pedraza J Kavanagh K Johannessen l Cuschieri K An evaluation of the Qiagen HPV sign for the detection and genotyping of cervical lesions and oropharyngeal squamous cell carcinomas J Virol Methods 2014 Oct 207 128 doi 101016 j jviromet 2014 07 0
6. the Scottish Cervical Screening Programme SCSP To this end SHPVRL currently performs HPV testing as a test of cure of treatment Following an early implementation phase across part of Scotland test of cure was rolled out Nationally from the 30th April in 2012 Procedures for the delivery of HPV testing services for test of cure have been developed between SHVPRL and the Scottish Cytology Laboratories for more detail please see section 6 page 9 SHPVRL collaborates with partners including UK NEGAS and the English Cervical Screening Programme in the preparation of material for HPV quality assurance and validation schemes Jof 28 Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 Review date 10 09 2016 e 3 CONTACT DETAILS AND HOURS OF WORK Name and Designation Director Dr Kate Cuschieri Medical Consultant Dr Ingo Johannessen Advanced Biomedical Scientist Mrs Catherine Moore Specialist Biomedical Scientist Miss Alison Fleming Associate Clinical Scientist Dr Christopher Ward Associate Clinical Scientist Mr Daniel Guerendiain Biomedical Support Worker Mr Norman Stenhouse SHPVRL Administrator Miss Kirsty Haxton Departmental confidential fax Royal Mail Postal Address Telephone numbers and Email address 0131 242 6039 Kate Cuschieri luht scot nhs uk 0131 242 6003 Ingo Johannessen luht scot nhs uk 0131 242 6005 Catherine Moore luht scot nhs uk 0131 242 6020 Alison Fl
7. 01 3 Cuschieri K Brewster DH Graham C Nicoll S Williams AR Murray GI Millan D Johannessen Hardie A Cubie HA Influence of HPV type on prognosis in patients diagnosed with invasive cervical cancer nt J Cancer 2014 Apr 17 doi 101002 ijc 28902 4 Kavanagh K Pollock KG Potts A Love J Cuschieri K Cubie H Robertson C Donaghy M Introduction and sustained high coverage of the HPV bivalent vaccine leads to a reduction in prevalence of HPV 16 18 and closely related HPV types Br J Cancer 2014 May 27 110 11 2804 11 doi 101038 bjc 2014 198 5 Cubie HA Canham M Moore C Pedraza J Graham C Cuschieri K Evaluation of commercial HPV assays in the context of post treatment follow up Scottish Test of Cure Study STOCS H J Clin Pathol 2014 Jun 67 6 458 63 doi 101138 jclinpath 2013 202014 Epub 2014 Jan 16 6 Cuschieri K Cubie H Graham C Rowan J Hardie A Horne A Earle CB Bailey A Crosbie EJ Kitchener H Clinical performance of RNA and DNA based HPV testing in a colposcopy setting Influence of assay target cut off and age J Clin Virol 2014 Feb 59 2 104 8 doi 10 1016 j jcv 2013 12 001 aj Jof 29 Author K Cuschieri Johannessen amp C Moore Doc no HPV 64 VERSION 4 Review date 10 09 2016 7 Cubie HA Cuschieri K Understanding HPV tests and their appropriate applications Cytopathology 2013 Oct 24 5 289 308 doi 101111 cyt12083 Epub 2013 Sep 2 Review PubMed PMID 23998275 8 Moor
8. CE marked and has been validated for genotyping of cervical liquid based cytology samples For biopsy samples the test used is the Optiplex HPV Genotyping Assay DiaMex which utilises luminex technology Detailed information on characteristics of tests and appropriate bio specimens for submission can be found in Tables 2 and 3 respectively For further information on HPV risk status please see Section 9 4 2 Assays used for Surveillance Testing SHPVHL performs HPV genotyping of biopsy material and cervical liquid based cytology samples in line with the National Surveillance System for HPV infection and related disease in Scotland d 15th October 2008 Genotyping results are generated using the Optiplex HPV Genotyping Assay Diamex Results are communicated directly to Health Protection Scotland via Electronic Communication of Surveillance information in Scotland ECOSS of 29 Author K Cuschieri Johannessen amp C Moore Doc no HPV 64 VERSION 4 Review date 10 09 2016 4 3 Other available HPV Assays SHPVRL has a role in evaluating the performance of new HPV tests with a mind to establishing which may be suitable for clinical use and or research and development going forward Having access to varied assays also enriches the annotation of quality materials Consequently several in house amp commercial HPV tests are available beyond those used for diagnostic testing and surveillance and include The Hybrid Cap
9. Doc no HPV 84 VERSION 4 Review date 10 09 2016 Scottish HPV Reference Laboratory SHPVRL Author K Cuschieri Johannessen amp C Moore NIS 3 a 1 P E t uncontrolled when printed _ ux Lothian M Doc no HPV 64 VERSION 4 Review date 10 09 2016 CONTENTS Glossary Page 3 Purpose Page 4 Scope of service Page 4 Contact details amp hours of work Page 5 HPV tests offered Page 6 Submission of specimens Page 8 Information for Scottish Cytology Labs Page 10 HPV Testing as a test of cure of treatment HPV types detected with different HPV tests Page 17 Factors affecting performance of HPV test Page 18 Additional tests Page 18 Definition of high risk and low risk HPV types Page 20 Reporting of results Page 23 Laboratory accreditation Page 23 External quality assurance schemes Page 23 SHPVRL and research and development Page 24 Selected publications Page 25 Appendix 1 Page 27 Appendix 2 Page 28 Appendix 3 Page 29 Page of 29 2 Author K Cuschieri Johannessen amp C Moore HPV HPS LBC SCSP ECOSS NATS SCCRS FFPE QCMD FDA CE IARC PV Doc no HPV 64 VERSION 4 Review date 10 09 2016 ID GLOSSARY Human Papillomavirus Health Protection Scotland Liquid Based Cytology Scottish Cervical Screening Programme Electronic Communication of Surveillance in Scotland Nucleic Acid tests Scottish Cervical Call Recall System Formali
10. Journal of Cancer 128 927 935 5 Schiffman et al 2009 Classification of weakly carcinogenic Human Papillomavirus types addressing the limits of epidemiology at the borderline Infectious Agents and Cancer 4 8 Page B of 29 Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 Review date 10 09 2016 de od eret ero 11 REPORTING OF RESULTS Written reports are issued and sent to the requesting consultant via the Royal Mail Emailed reports can be arranged provided the account is appropriate for secure transfer eg nhs net Transfer of surveillance results will be performed directly through ECOSS If requested results can be telephoned in advance of the report by a senior member of SHPVRL For advice and interpretation please contact Dr Kate Cuschieri or Dr Johannessen using contact details in Section HPV results associated with test of cure SCSP are disseminated through the national IT management tool for cervical screening the Scottish Cervical Call Recall System SCCRS At time or preparation of this manual we are working with SCSP Laboratory GA group to ensure that HPV results generated as a consequence of individual gynae clinical requests are entered SCCRS retrospectively ox wa ES 12 LABORATORY ACCREDITATION SHPVRL is situated within the Directorate of Laboratory Medicine Microbiology Laboratory which was accredited by CPA CPA Reference Number 2496 in May 2014 a
11. PS website and the RIE laboratory medicine website Request forms can be completed electronically or by hand Hard copies of request forms must accompany speciments www hps scot nhs uk reflab VirLabDetail aspx id2 26 www edinburghlabmed co uk Specialities reflab hpv Pages default aspx Where possible request forms should be printed from the websites above but can be provided by SHPVHL upon request Please use a separate request form for each patient ensuring that all relevant fields are completed As a minimum we require Patient s Identification Burname amp Forename CHI number Patient s date of birth and gender Specimen type site Consultant to whom a report should be addressed Location and contact details of sender Relevant clinical information Please note that if the sample is associated with the Scottish Cervical Screening programme test of cure service a form is not required as this workstream is organised through SCCRS please go to section 6 for details on test of cure processes 5 2 Specimen Labelling All specimens should be labelled with the following information Surname amp Forename CHI number for Scottish Referrals Date of birth A specimen and or associated request form with insufficient accompanying information as outlined above could result in a delay in the sample being tested or the sample not being tested p Jof es Author K Cuschieri Johannessen amp C Moo
12. been lysed with glacial acetic acid should be submitted for HPV testing however SHPVRL will be in touch with users either directly or via the cytology GA group with relevant updates It is essential that laboratories prepare specimens correctly to ensure accurate and timely result generation failure to do so will as a minimum delay result reporting When storing prequots prior to dispatch temperatures in excess of 25 C should be avoided Refrigerated 4 C storage of prequots is not mandatory but will not affect the test performance Prequots should not be frozen prior to transit Please ensure that caps are firmly secured given the methanol content in LBC samples minor leakage of a single sample within a batch can result in obliteration of the identifiers on all samples in the batch as well as posing a contamination risk Leaking samples are likely to result in delayed reporting or an inability to perform the assay Author K Cuschieri Johannessen amp C Moore Doc no HPV 64 VERSION 4 Review date 10 09 2016 SAMPLE ALIGUOTING FROM LBC VIALS Where possible aliquoting should be performed in a class 1 cabinet Howie coats and gloves should be worn and universal precautions adhered to Attach the relevant SCCRS label s to the orange capped tube s vertically The SCCHS label should be placed on the lower section of the tube with the left hand edge just above the skirt line See Figure 1 on page 12 Thoroughly mix the sampl
13. ction Proformas are provided by SHPVRL One copy of the proforma should be retained in the cytology laboratory attach to the SVG report list which should be filed in chronological order Contact courier to confirm date and time of the next uplift remember to include the date the samples were dispatched and the tracking number in the relevant section of the proforma For Lothian Cytology Laboratory Prequots should be stored in designated rack and transferred to SHPVRL Lab 55051 TURNAROUND FOR RESULT REPORTING For efficiency samples will be batched and SHPVRL will work to a turnaround of 5 working days The enquiries function in SCCRS will enable cytology staff to check the progress status of a particular sample However for specific enquiries please contact a member of SHPVHL using the details in section 3 page 4 41 Jof 29 Author K Cuschieri Johannessen amp C Moore OTHER QUESTIONS What are the time limits for test of cure HPV Testing Tests will not be accepted less than 4 months from treatment date but there is no upper time limit for inclusion in test of cure provided the patient has not previously been tested since treatment and there have been no further interventions What happens if a sample has been sent for test of cure in error Inform SHPVRL by phone it is possible to remove a sample from the virology module of SCCRS at any stage even if it has been tested The request for removal o
14. e C Cuschieri K McQueen F Duvall E Graham C Cubie HA Effect of glacial acetic acid pre treatment of cervical liquid based cytology specimens on the molecular detection of human papillomavirus Cytopathology 2013 Oct 24 5 314 20 doi 101111 cyt 12046 9 Junor E Kerr G Oniscu A Campbell 5 Kouzeli Gourley C Cuschieri Benefit of chemotherapy as part of treatment for HPV DNA positive but p16 negative squamous cell carcinoma of the oropharynx Br J Cancer 2012 Jan 17 106 2 358 65 doi 101038 bjc 2011 542 Author K Cuschieri Johannessen amp C Moore 1 Doc No HPV 53 Version 4 Review date 03 07 16 Gynaecology Form Human Papillomavirus HPV Test Request Form for GYNAE Scottish Human Papillomavirus Reference Laboratory Specialist Virology Centre Royal Infirmary of Edinburgh 51 Little France Crescent Edinburgh EH16 4SA From Sender Address PO Requestor Consultant Name Report will be sent to this person unless otherwise stated ee Address for Result Report if different from sender address PO Patient details Or place ID label that contains above info here Date amp time sample taken Hospital Laboratory ref no L Date posted nl Specimen LJ Cervical liquid based cytology sample LJ 2 x 10 um section of fixed biopsy i C Reason for request Referral from routine screening due to problems with cervical cytology oO Older 2 who have not e
15. e in the original LBC vial by by vortexing or by inverting the vial six times Using a pastette disposable plastic pipette remove sample from the LBC vial and expel into the corresponding orange tube The level of the liquid should be just over the upper right hand edge of the SCCRS label See Figure 1 this will constitute approximately 3 ml Dispose of the empty pastette Do not use a pastette for more than one sample as this will lead to cross contamination This is essential Ensure lid is FIRMLY secured store vial s in a designated area pending dispatch to SHPVRL External cytology labs are expected to purchase the prequot tubes updated supplier information is below Company Abbott Molecular Product number 4J7180 Product Description Mastermix Tubes Unit Pack of 150 Author K Cuschieri Johannessen amp C Moore Doc no HPV 64 VERSION 4 Review date 10 09 2016 Figure 1 Mock up of an appropriately labeled Prequot Lid firmly secured Using a pastette add sample to a level equal to or just over the right hand edge barcode label placed vertically on side of tube Place SCCRs barcode label vertically on side of tube with the left hand edge just above the skirt line MANDATORY DATA SET All prequots samples must be labeled with the appropriate SCCRS label this will contain SCCRS no Cytology Lab Accession no Patient Name CHI Failure to label the prequots will make them totally unid
16. eming luht scot nhs uk 0131 242 6020 ChristopherWard nhslothian scot nhs uk 0131 242 6020 Daniel Guerendiain nhslothian scot nhs uk 0131 242 6020 Norman Stenhouse luht scot nhs uk 0131 242 6010 Kirsty Haxton nhslothian scot nhs uk 0131 242 6008 Scottish Human Papillomavirus Reference Laboratory SHPVRL Division of Laboratory Medicine Microbiology Department Edinburgh Royal Infirmary 51 Little France Crescent Edinburgh EH16 4SA Hays DX Address number 6231202 Exchange depot Edinburgh 96 EH Hours of work Core laboratory hours are 9am 5pm Monday to Friday SHPVRL does not currently offer an out of hours service Jof 28 Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 Review date 10 09 2016 4 HPV TESTS OFFERED 44 For diagnostic testing SHPVRL can perform HPV screening and or genotyping where an HPV result would inform and support individual patient management All detection assays used currently at SHPVRL are nucleic acid tests NATS see section 7 Table 2 Current assays used for clinical testing are For high risk HPV detection consensus testing of a group of high risk types RealTime HPV Assay Abbott This assay is CE marked and is one of the HPV tests that have been approved for use within the English Cervical Screening Programme For type specific genotyping Roche linear array HPV Genotyping Test Roche Molecular Systems is used This assay is
17. entifiable so it is essential that correct labels be attached SHPVRL require an additional aliquot Occasionally virology will Require additional aliquot from cytology and this will appear as an alert on SCCRS this is not a definitive result and Cytology will be tasked with providing and sending a repeat aliquot of the specimen if available 5 Jot 29 Author K Cuschieri Johannessen amp C Moore Doc no HPV 64 VERSION 4 Review date 10 09 2016 SAMPLE TRANSFER to SHPVRL non Lothian Prequots for test of cure should be batched where possible Standard Hayes container will take around 12 orange capped tubes to ensure efficiency of transfers Tubes should be placed in Hayes DX container or approved alternative according to packaging instructions Packaging should be in accordance with UN 3373 Biological Substance category B transport regulations and conform to packing instructions 650 PI650 When collating a batch of aliquots for transportation to SHPVRL the individual responsible for this task must check the SVG to ensure there is tubes for all selected samples and there are no duplicate or inappropriate extra tubes for samples not on the SVG For each transfer the proforma Specimen transfer from Oytology for Test of Cure HPV testing should be completed and dispatched with the samples in between the biobottle and the cardboard outer Please complete all fields other than those within For SHPVRL use only se
18. f a sample will appear on the audit trail What happens if virology have amended updated an HPV test result If virology have updated an HPV test result an alert will appear on the Labs Home page of SCCRS The amended virology result will automatically update the CCR final result screen and this will appear on the audit trail What happens if a sample that should have had an HPV test as a test of cure is missed and the result has been reported in cytology For this scenario if cytology laboratory still has the relevant sample an aliquot can be sent with a clinical request form see appendix please write clearly in the reason for request section of the form that the sample relates to a missed test of cure What happens if the sample is rejected by virology SHPVRL SHPVRL will inform the cytology lab if a sample has been rejected reasons for this include insufficient volume or leakage within the transit container A further aliquot should be provided if available If it is not possible to provide a further aliquot inform SHPVRL and de select sample from the SCCRS HPV queue Author K Cuschieri Johannessen amp C Moore Can we request additional investigations on samples Other than repeating the Abbott assay where required necessary e g in certain cases where an additional aliquot is required additional HPV tests are not offered as part of Test of Cure If SHPVRL require an additional aliquot an alert wi
19. ing There is no defined time limit for requesting additional examinations but if the request is possible their performance may affect the stated turnaround table 2 Turnaround is based on when the specimen request was initially received All additional assays whether generated in house or requested by users will be adjudicated on a case by case basis by a senior member of SHPVRL Jof 29 Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 Review date 10 09 2016 10 DEFINITION OF HIGH RISK AND LOW RISK HPV TYPES 10 1 Overview of classification HPV types are often categorised as high or low risk according to the strength of their association with cancer All HPV genotypes that are known to be cervical carcinogens belong to the alpha genus Within the alpha genus species groups which unite related types which have been found to contain high risk types are alpha 5 alpha 6 alpha 7 alpha 9 and alpha 11 The evidence base which has informed the risk classifications is drawn from worldwide surveys of HPV type distribution in high grade cervical lesions and cancers This evidence base continues to accrue and as a consequence the classifications refine In addition attribution of risk for rare types is challenging 10 2 Categorisation of HPV types according to standardised grouping system for human carcinogens In 2009 an expert working group at the International Agency fo
20. ll appear on SCCRS Require additional aliquot for SCCRS No etc If there is no or insufficient material to send a repeat aliquot please inform SHPVRL who will update the HPV status of the result to Fail Will the HPV surveillance process be affected by test of cure The cytology laboratory collection processes for National HPV surveillance where LBC samples from the screening population are collected over a fixed period and sent to SHPVRL should not affect or interfere with any of the processes for test of cure However for any queries relating to this please contact SHPVRL Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 Review date 10 09 2016 7 HPV TYPES DETECTED WITH DIFFERENT HPV TESTS Different commercial HPV tests are designed to detect a specific range of HPV types dependent on the technology used and the primer probe range incorporated These are detailed in Table 2 Table 2 Type ranges of HPV tests performed High risk HPV detection rT HPV Assay Abbott This Min 2 x Weekly 5 working days consensus testing cervical is a CE marked assay which clinical requests LBC samples detects 14 high risk HPV 5 working days test types in aggregate and of provides typing information on presence absence of HPV 16 and 18 HPV type specific Linear Array HPV Genotyping Weekly to fortnightly 10 days genotyping Cervical LBC test Roche depending on number of samples reques
21. m for non GYNAE Scottish Human Papillomavirus Reference Laboratory Specialist Virology Centre Royal Infirmary of Edinburgh 51 Little France Crescent Edinburgh EH16 4SA From Sender Address DOO f Requestor Consultant Name Report will be sent to this person unless otherwise stated PO Address for Result Report if different from sender address Patient details Hospital Laboratory ref no Date of birth O Date amp time sample taken of E Date posted Specimen Test required 1 2 x 10 um section of fixed biopsy High risk HPV genotyping LJ Original block High risk HPV genotyping Is this an Oropharyngeal Biopsy _ Yes No If not an Oropharyngeal Biopsy specify site Please note that biopsies from outside the oropharynx should p16 Status of specimen be discussed with SHPVRL prior to submission L Pos Unclear Not done If p16 staining is to be performed please let u Will p16 Staining be performed Yes LINO SHPVRL know the result when available Reason for request All cases must be discussed at multi disciplinary team meeting For further information please contact Dr Kate Cuschieri on 0131 242 6039 or Dr Ingo Johannessen on 0131 242 6003 SHPVRL LAB USE ONLY Comments Test code Lo fL Page of 29 Authority for issue K Cuschieri Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 e APPENDIX
22. n Fixed Paraffin Embedded National External Quality Assurance Scheme Quality Control in Molecular Diagnostics Food and Drug Administration Certificate European International Agency of Research on Cancer Papillomavirus Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 Review date 10 09 2016 e 1 PURPOSE The Scottish Human Papillomavirus Reference Laboratory SHPVRL is commissioned by Health Protection Scotland HPS to provide a comprehensive service for screening and typing of Human Papillomavirus HPV e 2 SCOPE OF SERVICE To provide testing services to evaluate the impact of the HPV immunisation programme on the incidence and prevalence of HPV related disease and HPV infection in the Scottish population To provide a specialist diagnostic and expert advice service in relation to HPV for specific clinical cases where it could help inform clinical management To provide education and training for the wider service and develop and validate new HPV technologies carry out quality control and external quality assurance and act as a base for research and development To contribute to the data and knowledge of changing epidemiology of HPV associated disease to help inform the future organisation of services to prevent cervical disease and cancer Other elements HPV testing for the Scottish Cervical Screening Programme SCSP SHPVRL also provides information advice and testing services for
23. nd found to be in conformance with the Clinical Pathology Accreditation UK Ltd Standards for the Medical Laboratory gt TORO EJ 13 EXTERNAL QUALITY ASSURANCE SCHEMES National External Quality Assurance NEGAS SHPVRL takes part in the HPV UK NEGAS scheme this involves the testing of a panel of 4 unknown clinical samples distributed three times per year The scheme interrogates the screening performance of assays and is designed to support clinical testing Quality Control for Molecular Diagnostics GCMD SHPVRL takes part in the HPV GCMD scheme which involves the testing of a panel of 10 samples distributed once per year The scheme interrogates the screening performance of assays WHO HPV LabNet SHPVRL take part in the WHO LabNet EGA scheme The scheme interrogates the type specific performance of assays 5 Jof 29 Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 Review date 10 09 2016 14 SHPVRL AND RESEARCH AND DEVELOPMENT SHPVRL has a keen interest and commitment to research and development Particular areas of interest are Working with public health the cervical screening programme and academia to ensure that optimal systems for both HPV surveillance and cervical disease management and prevention are available e Assay evaluation and development in conjunction with NHS academic and industrial partners e Acting as clinical laboratory partners for external basic
24. r Research on Cancer IARC endeavoured to categorise HPV types according to a standardised grouping system for human carcinogens thus Group 1 Carcinogenic to humans Group 2A Probably carcinogenic to humans Group 2B Possibly carcinogenic to humans Group 3 Not classifiable as to its carcinogenicity in humans Table 3 shows the HPV types incorporated into the genotyping assays used at SHPVRL their associated carcinogenic group according to IARC and risk status according to current evidence expert opinion of 29 Author K Cuschieri Johannessen amp C Moore Doc no HPV 64 VERSION 4 Review date 10 09 2016 Table Carcinogenic potential risk status of HPV types detected in assays used at 11 16 18 26 31 33 35 39 40 42 43 44 45 51 52 53 54 55 56 58 59 61 62 64 66 67 SHPVRL 10 10 gt N D 0 E o 510 pals o O O A Unlikely Low risk Unlikely Low risk Yes High risk Yes High risk Possibly Probably high risk or query high risk Yes High risk Yes High risk Yes High risk Yes High risk Unlikely Low risk Unlikely Low risk Unlikely Low risk Unlikely Low risk Yes High risk Yes High risk Yes High risk Possibly Probably high risk or query high risk Unlikely Low risk Unlikely Low risk Yes High risk Yes High risk Yes High risk Unlikely Low risk Unlikely Low risk Possibly Probably high risk or query high risk Possibly Probably high ri
25. re 5 3 Courier transit instructions All specimens should be sent in accordance with UN 3373 Biological Substance category B transport regulations and conform to packing instructions 650 650 The courier HAYS DX can be used to transport samples to SHPVHL see section 3 Doc no HPV 84 VERSION 4 Review date 10 09 2016 For users sending samples accompanied with a sample receipt form that requires acknowledgement SHPVRL will not confirm receipt by fax but by phone date of call will be logged for audit purposes 5 4 Specimen types appropriate for clinical requests Table 1 Appropriate specimens for diagnostic HPV testing High risk HPV detection consensus testing or screening HPV genotyping type specific detection Other types of samples can only be acce Cervical Liquid Based Cytology LBC sample in PreservCyt Media in original LBC container OR A 3 ml prequot can be removed The term prequot refers to an aliquot that is removed from the original LBC vial before it is transferred to an automated slide processor For biopsies 2 10 um sections of Formalin Fixed Paraffin Embedded Material placed in separate sterile 2 ml Screw cap tubes ie maximum of 1 10 um section per tube OR Original relevant FFPE block on which pathology diagnosis has been made in sealed labeled plastic bag Cervical Liquid Based Cytology sample in PreservCyt Media in original LBC container OR 3 ml prequot
26. rol for sample quality Thus if a sample tests HPV negative and is also negative for the human DNA gene a result of invalid is generated The RealTime HPV test also incorporates a human cellular control For laboratories sending paraffin sections it is essential to refer to the protocol for sectioning paraffin blocks at source laboratory See Appendix 3 As the downstream HPV test used for FFPE sections is very sensitive we therefore ask laboratories to send a section from a blank block if sending a section for diagnostic HPV testing to SHPVRL for the first time to monitor any potential environmental contamination Testing of blank sections is monitored to inform frequency of additional requests for blank sections Jof 29 Author K Cuschieri Johannessen amp C Moore Doc no HPV 64 VERSION 4 Review date 10 09 2016 ADDITIONAL TESTS SHPVRL decision SHPVRL may perform a confirmatory HPV test on a specimen beyond that requested by the user if we consider that it would aid interpretation and be of benefit to the user An example would be performing an HPV genotyping test on a sample we received for high risk HPV screening which screened positive For these cases results would be aggregated in one report User requests For users who wish to organise an additional examination on a sample these may may not be possible depending on the nature of the request and the amount of residual specimen remain
27. sk or query high risk Possibly Probably high risk or query high risk Page Jof 29 Most potent type Risk of cancer an order of magnitude higher compared to other Group 1 high risk types Subtype of HPV 44 Subtype of 34 Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 Review date 10 09 2018 Table 3 Continued Probably High risk 69 5 Possibly Probably high risk or query high risk 70 7i Possibly Probably high risk or query high risk 71 14 Unlikely Low risk 78 3 Unlikely Low risk 73 11 Possibly Probably high risk or query high risk 74 10 Unlikely Low risk 81 3 Unlikely Low risk 82 5 Possibly Probably high risk or query high risk 83 3 Unlikely Low risk 84 3 Unlikely Low risk HPVIS 5 Possibly Probably high risk or Subtype of HPV 82 139 query high risk HPV 3 Unlikely Low risk or query risk Candidate HPV 89 CP6108 The table has been constructed from the following sources 1 Bouvard V et al 2009 Special Report Policy A review of human carcinogens part B biological agents 2 De Villiers et al 2004 Classification of papillomavirus Virology 324 17 27 3 Bernard et al 2010 Classification of papillomaviruses based on 189 PV types and proposal of taxonomic amendments Virology 401 70 79 4 2011 Human Papillomavirus type distribution in 30 848 invasive cervical cancers worldwide variation by geographical region histological type and year of publication International
28. ts HPV type specific genotyping Optiplex Multiplex HPV Weekly 10 days Biopsy Samples LBC Genotyping Kit Diamex samples for surveillance Turnaround time excludes weekends and public holidays a Detection Range 16 18 31 33 35 39 45 51 52 56 58 59 66 68 b Detection range HPV 6111618 26 31 33 35 39 404245 51 52 53 54 55 56 58 59 61 62 64 66 6768 69 70 71702 73 8182 83 84 HPVIS39 HPV CP6108 c Detection range 6111618 26 3133 35 39 42 43 44 45 51 52 53 56 58 59 66 68 70 73 82 For risk status associated with the types please see section 9 Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 Review date 10 09 2016 8 FACTORS AFFECTING PERFORMANCE OF HPV TEST The yield and quality of nucleic acid in cervical liquid based cytology samples collected in PreservCyt is high However when storing liquid based cytology samples prior to dispatch temperatures in excess of 25 C should be avoided Refrigerated 4 C storage of prequots is not mandatory but will not affect the test performance Liquid based cytology samples should not be frozen prior to transit Degradation fragmentation of nucleic acid can however impair the performance of the HPV NATs Degradation is more associated with paraffin embedded material than with liquid based cytology samples particularly where these have been stored in excess of 2 years The genotyping assays used by SHPVRL incorporate amplification of a human DNA gene to cont
29. ture 2 assay Giagen a consensus DNA based screening assay Consensus mRNA based screening assays and type specific mRNA assays are also available All new tests are subjected to validation of the technology evaluation of performance and effectiveness appropriate to the work stream diagnostic research epidemiology Users would be informed of any changes made in tests relating to service in advance of the next version of this user manual Costs associated with HPV testing For NHS Boards in Scotland Finite funding has been allocated for diagnostic HPV tests relating to requests from NHS Boards in Scotland Thus tests are free for referrals from NHS Boards in Scotland until the allocation is fully utilised For NHS Boards outwith Scotland and for Private Institutions Costs have been calculated for non Scottish NHS Boards and for private requests Hequests for undertaking HPV testing in these situations must be discussed with SHPVRL before sending specimens Jot 89 Author K Cuschieri Johannessen amp C Moore Doc no HPV 84 VERSION 4 Review date 10 09 2016 5 SUBMISSION OF SPECIMENS Individual Clinical Requests 5 1 Request form All specimens must be accompanied with the appropriate request form Please complete the relevant SHPVRL request form for either gynaecological or non gynaecological requests Both forms can be found in appendix 2 and 3 of this user manual and are also included as separate PDFs on the H
30. xited the SCSP because of continuing low grade LG disease Patients where there is difficulty obtaining liquid based cytology sample e g Immunosuppresion Al disease Exceptional cases not included here where knowledge of HPV status will influence management these cases must be discussed at Colposcopy MDT Details e i Test required LI High risk HPV screening HPV Genotyping Please note that other biospecimen types can only be accepted after prior discussion with SHPVRL Referral following problems associated with colposcopy visits Persistent LG abnormality after treatment where fertility conservation is an issue Postmenopausal with persistent LG abnormalities attending Colposcopy given HRT if colp inadequate difficulty identifying SCJ where LETZ shown no histological abnormality amp vaginal colposcopic assessment is negative HIV ve patients with persistent LG abnormalities where knowledge of HR HPV ve status would allow expectant observational management Discussed E Dale For further information please contact Dr Kate Cuschieri 0131 242 6039 or Dr Ingo Johannessen on 0131 242 6003 SHPVRL LAB USE ONLY Comments Authority for issue K Cuschieri Page Test code s of 29 Author K Cuschieri Johannessen amp C Moore e aa Doc No HPV 54 Version 4 Review date 03 07 16 Non Gynae Request Form Human Papilloma Virus HPV Test Request For
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