WHO Prequalification Of In Vitro Diagnostics Programme .
PQDx 0236-078-00WHO PQ Public ReportOctober 2020, version 6.0WHO Prequalification of In Vitro Diagnostics ProgrammePUBLIC REPORTProduct: Aptima HIV-1 Quant Dx AssayWHO Reference number: PQDx 0236-078-00Aptima HIV-1 Quant Dx Assay with product codes PRD-03000 (PRD-03002, PRD-03001),303014 and PRD-03003, and instrument Panther system with product code 303095,manufactured by Hologic, Inc, CE regulatory version, was accepted for the WHO list ofprequalified in vitro diagnostics and was listed on 21 December 2017.Summary of WHO prequalification assessmentfor Aptima HIV-1 Quant Dx AssayPrequalification listingDossier assessmentSite inspection(s) of quality managementsystemProduct performance evaluationDateOutcome21-Dec-2017 listedN/A MR1 to 2-Jul-2015 MR14-Dec-2017 MRMR: Meets RequirementsNA: Not ApplicableReport amendments and/or product changesThis public report has since been amended. Amendments may have arisen because ofchanges to the prequalified product for which WHO has been notified and has undertakena review. Amendments to the report are summarized in the following table, and details ofeach amendment are provided below.Version Summary of amendment1.0-4.05.0Date of reportamendmentEditorial changes to the initial public report for listing.21-Dec-20171. Transfer of assay reagents and equipment used to manufacture 07-Feb-2018Aptima HIV-1 Quant Dx Assay from the Willow Court facility to theGenetic Center Drive facility.2. Labelling change to include additional print on the existing labellingstating the new manufacturing site.Page 1 of 94
PQDx 0236-078-006.0WHO PQ Public ReportOctober 2020, version 6.0Adding Dried Blood Spots (DBS) sample type to the Aptima HIV-1 Quant 27-Oct-2019Dx Assay. Labelling change to include additional print on the finishedlabel of the Aptima DBS Extraction Buffer which states the Manchestermanufacturing address.Intended use:According to the claim of intended use from Hologic, Inc., for the Aptima HIV-1 Quant Dxassay package insert “the Aptima HIV-1 Quant Dx and DBS Supplement to the Aptima HIV-1Quant Dx Assay is an in vitro nucleic acid amplification test for the detection and quantitationof human immunodeficiency virus type 1 (HIV-1) RNA groups M, N, and O on the fullyautomated Panther system. It is intended for use as an aid in the diagnosis of HIV-1infection, as a confirmation of HIV-1 infection, and as an aid in clinical management ofpatients infected with HIV-1.The Aptima HIV-1 Quant Dx assay may be used as an aid in the diagnosis of HIV-1 infection,including acute or primary infection. Presence of HIV-1 RNA in the plasma or serum ofpatients without antibodies to HIV-1 is indicative of acute or primary HIV-1 infection.The Aptima HIV-1 Quant Dx assay may be used as a supplemental test for specimens thathave repeat reactive results with approved HIV immunoassays. If the specimen is reactive inthe Aptima HIV-1 Quant Dx assay, HIV-1 infection is confirmed.The Aptima HIV-1 Quant Dx assay may also be used in conjunction with clinical presentationand other laboratory markers for disease prognosis in HIV-1 infected individuals. The AptimaHIV-1 Quant Dx assay may be used as an aid in monitoring the effect of antiretroviraltreatment by measuring changes in the concentration of HIV-1 RNA in plasma.When the Aptima HIV-1 Quant Dx assay is used as an aid in the diagnosis of HIV-1 infection,performance for qualitative results is established with both plasma and serum specimens. *When used as an aid in monitoring the effect of antiretroviral therapy, performance forquantitative results is established with plasma specimens only. Serum specimens may not beused for quantitative results.This assay is not intended for use in screening blood or plasma donors.”According to the claim of intended use from Hologic, Inc., DBS Supplement to the AptimaHIV-1 Quant Dx Assay, “the Aptima HIV-1 Quant Dx assay is an in vitro nucleic acidamplification test for the detection and quantitation of human immunodeficiency virus type1 (HIV-1) RNA groups M, N, and O on the fully automated Panther system. It is intended foruse as an aid in the diagnosis of HIV-1 infection, as a confirmation of HIV-1 infection, and asan aid in clinical management of patients infected with HIV-1.Page 2 of 94
PQDx 0236-078-00WHO PQ Public ReportOctober 2020, version 6.0In addition, the Aptima HIV-1 Quant Dx assay may be used as an aid in the diagnosis ofacute or primary HIV-1 infection. Presence of HIV-1 RNA in the plasma, serum, or blood ofpatients without antibodies to HIV-1 is indicative of acute or primary HIV-1 infection. TheAptima HIV-1 Quant Dx assay may be used as a supplemental test for specimens that haverepeat reactive results with approved HIV immunoassays. If the specimen is reactive in theAptima HIV-1 Quant Dx assay, HIV-1 infection is confirmed.The Aptima HIV-1 Quant Dx assay may also be used in conjunction with clinical presentationand other laboratory markers for disease prognosis in HIV-1 infected individuals. The AptimaHIV-1 Quant Dx assay may also be used as an aid in EID of HIV-1 infection in infants below18 months of age using DBS. The Aptima HIV-1 Quant Dx assay may be used as an aid inmonitoring the effect of antiretroviral treatment by measuring changes in the concentrationof HIV-1 RNA in plasma and DBS samples.When the Aptima HIV-1 Quant Dx assay is used as an aid in the diagnosis of HIV-1infection, performance for qualitative results is established with both plasma and serumspecimens as well as DBS samples from infants below 18 months of age. When used as anaid in monitoring the effect of antiretroviral therapy, performance for quantitative results isestablished with plasma and DBS specimens only. Serum specimens may not be used forquantitative results.This assay is not intended for use in screening blood or plasma donors.”Assay description:According to the claim of intended use from Hologic, Inc, “Aptima HIV-1 Quant Dx assayinvolves three main steps, which all take place in a single tube on the Panther system: targetcapture, target amplification by transcription-mediated amplification (TMA), and detectionof the amplification products (amplicon) by the fluorescent labeled probes (torches).During target capture, viral nucleic acids are isolated from specimens. The specimen istreated with a detergent to solubilize the viral envelope, denature proteins, and release viralgenomic RNA. Capture oligonucleotides hybridize to highly conserved regions of the HIV-1genome, if present, in the test specimen. The hybridized target is then captured ontomagnetic microparticles that are separated from the specimen in a magnetic field. Washsteps remove extraneous components from the reaction tube.Target amplification occurs via TMA, which is a transcription-mediated nucleic acidamplification method that utilizes two enzymes, MMLV (Moloney murine leukemia virus)reverse transcriptase and T7 RNA polymerase. The reverse transcriptase is used to generatea DNA copy (containing a promoter sequence for T7 RNA polymerase) of the target sequence.T7 RNA polymerase produces multiple copies of RNA amplicon from the DNA copy template.The Aptima HIV-1 Quant Dx assay utilizes the TMA method to amplify two regions of HIV-1Page 3 of 94
PQDx 0236-078-00WHO PQ Public ReportOctober 2020, version 6.0RNA (pol and LTR). Amplification of these specific regions is achieved using specific primerswhich are designed to amplify HIV-1 groups M, N, and O. The primer design and the dualtarget approach ensure accurate detection and quantitation of HIV-1.Detection is achieved using single-stranded nucleic acid torches that are present during theamplification of the target and that hybridize specifically to the amplicon in real-time. Eachtorch has a fluorophore and a quencher. When the torch is not hybridized to the amplicon,the quencher is in close proximity of the fluorophore and suppresses the fluorescence. Whenthe torch binds to the amplicon, the quencher is moved farther away from the fluorophoreand it will emit a signal at a specific wavelength when excited by a light source. As moretorches hybridize to amplicon a higher fluorescent signal is generated. The time taken for thefluorescent signal to reach a specified threshold is proportional to the starting HIV-1concentration. Each reaction has an internal calibrator/internal control (IC) that controls forvariations in specimen processing, amplification, and detection. The concentration of asample is determined by the Panther system software using the HIV-1 and IC signals for eachreaction and comparing them to calibration information.”Test kit contents:Aptima HIV-1 Quant Dx Assay Box (Product code PRD-03000)ComponentA qHIV-1 AmplificationReagentE qHIV-1 Enzyme ReagentPRO qHIV-1 PromoterReagentAR qHIV-1 AmplificationReconstitution SolutionER qHIV-1 EnzymeReconstitution SolutionPR qHIV-1 PromoterReconstitution SolutionTCR qHIV-1 Target CaptureReagentDescriptionNon-infectious nucleicacids dried in bufferedsolution.Reverse transcriptase andRNA polymerase dried inHEPES bufferedNon-infectious nucleicacids dried in bufferedsolution.Aqueous solutioncontaining glycerol andpreservativesHEPES buffered solutioncontaining a surfactantand glycerolAqueous solutioncontaining glycerol andpreservatives.Nucleic acids in a bufferedsalt solution containingsolid phase, noninfectiousPage 4 of 94Quantity (for 100 tests)1 vial (lyophilized)1 vial (lyophilized)1 vial (lyophilized)1 vial x 7.2 ml1 vial x 5.8 ml1 vial x 4.5 ml1 vial x 72.0 ml
PQDx 0236-078-00Reconstitution CollarsMaster Lot Barcode SheetWHO PQ Public ReportOctober 2020, version 6.0nucleic acids, and InternalCalibrator.N/AN/A31 sheetNote: PRD-03000 contains 1 assay box, 1 calibrator kit, and 1 controls kit. Calibrator andControl kits are available separately if desired.Aptima HIV-1 Quant Dx Controls Kit (Product code PRD-03002)ComponentNC qHIV-1 Negative ControlLPC qHIV-1 Low PositiveControlHPC qHIV-1 High PositiveControlControl Barcode LabelDescriptionHIV-1 negativedefibrinated humanplasma containinggentamicin and0.2% sodium azide aspreservatives.Non-infectious HIV-1Armored RNA indefibrinated humanplasma containinggentamicin and 0.2%sodium azide aspreservatives.Non-infectious HIV-1Armored RNA indefibrinated humanplasma containinggentamicin and 0.2%sodium azide aspreservatives.Quantity5 vials x 1.5 ml5 vials x 1.5 ml5 vials x 1.5 ml-Aptima HIV-1 Quant Dx Calibrator Kit (Product code PRD-03001)ComponentPCAL qHIV-1 PositiveCalibratorCalibrator Barcode LabelDescriptionTranscript in bufferedsolution.Page 5 of 94Quantity (for 100 tests)5 vials x 2.5 ml-
PQDx 0236-078-00WHO PQ Public ReportOctober 2020, version 6.0Aptima DBS Extraction Buffer (Product code: PRD-04772).ComponentDBS extraction bufferDescriptionPhosphate bufferedsolution containingdetergentQuantity (for 100 tests)1 vial x 104 mlItems required but not provided, available separately.Note: Materials available from Hologic have product codes listed, unless otherwisespecified.ComponentInstrumentPanther systemConsumablesPanther Run Kit for Real Time AssaysProduct codeDescription303095InstrumentPRD-03455(5000 tests)Aptima Assay Fluids kit (Universal Fluids kit)Aptima Specimen DiluentMulti-tube units (MTUs)Panther Waste Bag KitPanther Waste Bin CoverTips, 1000 μL conductive, liquid sensing, TecanBleach, 5% to 7% (0.7M to 1.0M) sodiumhypochlorite solutionDisposable, powderless glovesReplacement non-penetrable capsReagent replacement caps: Amplification, Enzyme, and Promoterreagent reconstitution bottles (100caps) TCR bottle (100 caps)Assay Fluids kitMulti-tube UnitsPanther waste bagkitPanther waste bincover303014 (1000 Aptima Washtests)Solution, AptimaBuffer forDeactivation Fluid,andAptima Oil ReagentPRD 03036ACL0041CL0040Page 6 of 94
PQDx 0236-078-00WHO PQ Public ReportPlastic-backed laboratory bench coversLint-free wipesPipettorTipsPrimary collection tubes (ACD, EDTA, PPT, SST,Serum) of the following dimensions may beused: 13 mm x 100 mm 13 mm x 75 mm 16 mm x 100 mmDBS Extraction materials.Aptima specimen aliquot tubesTransport tube capsCalibrated PipettorsAerosol barrier pipette tipsEquipmentCentrifugeVortex mixerOctober 2020, version 6.0-503762504415-Pack of 100Pack of 100--Storage:Aptima HIV-1 Quant Dx Assay Box should be stored at 2-8 C.Aptima HIV-1 Quant Dx Calibrator Kit should be stored at -15 C to -35 C.Aptima HIV-1 Quant Dx Controls Kit should be stored at -15 C to -35 C.Aptima DBS Extraction Buffer should be stored at 15 C to 40 C.Shelf-life upon manufacture:24 months.Warnings/Limitations:Refer to the attached instructions of use (IFU).Prioritization for prequalificationBased on the established eligibility criteria, Aptima HIV-1 Quant Dx Assay was given priorityfor WHO prequalification assessment.Page 7 of 94
PQDx 0236-078-00WHO PQ Public ReportOctober 2020, version 6.0Product dossier assessmentIn accordance with the WHO procedure for abbreviated prequalification assessment,Hologic Inc. was not required to submit a product dossier for Aptima HIV-1 Quant Dx Assayas per the “Instructions for compilation of a product dossier” (PQDx 018 v1).Notwithstanding, certain aspects of the product dossier previously submitted for stringentregulatory review were reviewed by an assessor during the site inspection.Commitment for prequalificationAs a commitment for prequalification, the manufacturer committed to amend the IFU toinclude a warning that only EDTA plasma has been validated by 30 April 2021.Manufacturing site inspectionIn accordance with the WHO procedure for abbreviated prequalification assessment, ashortened inspection with fewer inspectors was conducted at the site(s) of manufacture(10210 Genetic Center Drive, San Diego, 92121, USA, 6333 Sequence Drive, San Diego,92121 USA, and 10808 Willow Court, San Diego, 92127, USA) of Aptima HIV-1 Quant DxAssay in July 2015 per the “Information for manufacturers on prequalification inspectionprocedures for the sites of manufacture of diagnostics” (PQDx 014 v1).The inspection found that the manufacturer had an acceptable quality management systemand good manufacturing practices in place that ensured the consistent manufacture of aproduct of good quality.The manufacturer's responses to the nonconformities found at the time of the inspectionwere accepted 6 August 2015.Commitment for prequalification:Include WHO in the reporting system to ensure information is communicated to WHO onany adverse event or safety corrective action.Page 8 of 94
PQDx 0236-078-00WHO PQ Public ReportOctober 2020, version 6.0Product performance evaluationAptima HIV-1 Quant Dx Assay was evaluated from 28 October 2015 to 6 April 2016 and from29 November 2016 to 23 July 2017. From this evaluation, we drew the following conclusions.Aptima HIV-1 Quant Dx Assay is a laboratory-based in vitro nucleic acid amplification testfor the detection and quantitation of human immunodeficiency virus type 1 (HIV-1) RNAgroups M, N, and O on the fully automated Panther system. It is intended for use as an aidin the diagnosis of HIV-1 infection, as a confirmation of HIV-1 infection, and as an aid inclinical management of patients infected with HIV-1.This type of assay requires additional laboratory equipment for specimen preparation andreagent storage and cannot be performed in laboratories with limited facilities. Theinstrument requires a stable source of electricity and has a one meter square footprint.Furthermore, substantial training was considered essential to obtaining accurate results.Analytical specimens:The assay’s precision of measurement was verified. In this evaluation the precision ofmeasurement was found to be acceptable, all %CV were found to be 6%.The linearity of the assay was verified in Subtypes A, B, C, D, and AG. In this evaluation thelinearity for all subtypes were estimated by linear regression. All slopes were 0.2 from anideal value of 1. R2 values were all 0.97 indicating good correlation between the referencemethod and the assay under evaluationThe limit of detection was verified. In this evaluation the LOD was estimated to be 31.57IU/ml (95% Fiducial limits: 20.63-75.37); 11.05 copies/ml (95% Fiducial limits: 7.22-26.38,conversion factor is 0.35, 1 IU 0.35 copies).No carry-over was detected.Clinical specimens – first evaluation:In this performance evaluation on a panel of 432 specimens, we found a bias -0.065 log10copies/ml (limits of agreement: -1.158; 1.027) compared to the reference results.Correlation was found to be within range (R2 0.9476, P 0.1).Sensitivity for treatment failure at 1000 copies/ml 93.91% (95% CI: 90.00-96.34)Specificity for treatment failure at 1000 copies/ml was 100.00% (95% CI: 98.13-100.00)In this study, the invalid rate was 3.3%.Page 9 of 94
PQDx 0236-078-00WHO PQ Public ReportPerformance characteristicsAnalytical performanceLimit of DetectionLinearityCarry-overClinical performanceBiasOctober 2020, version 6.031.57 IU/ml (95% Fiducial limits: 20.63-75.37);11.05 copies/ml (95% Fiducial limits: 7.22-26.38)Verified in subtypes: A, B, C, D, and AGLinearity for all subtypes was determined to beacceptable. All slopes were within were 0.2 froman ideal value of 1. R2 values were all 0.970%-0.065 log10 copies/ml (limits of agreement: -1.158;1.027)CorrelationWithin range (R2 0.9476, P 0.1)Sensitivity for virological failure at 93.91% (95% CI: 90.00-96.34)1000 copies/mlSpecificity for virological failure at 100.00% (95% CI: 98.13-100.00)1000 copies/mlInvalid rate3.29%Key operational characteristicsValidated specimen typesNumber of stepsTime to resultIn-use stability of reagentsFor quantitative measurements:Tubes containing EDTA or Acid Citrate Dextrose(ACD) anticoagulants orPlasma Preparation Tubes (PPTs).For qualitative determination:Tubes containing EDTA or ACD anticoagulants, orPPTs, orSerum tubes, orSerum separator tubes (SSTs).103h:30 minutes (preparation and loading: 50 minutes;test: 2h:40 minutes for the first result; 5 resultsevery 5 minutes thereafter).30 days for reagents20 hours for single use vialsPage 10 of 94
PQDx 0236-078-00WHO PQ Public ReportLabelling1.Labels2.Instructions for usePage 11 of 94October 2020, version 6.0
PQDx 0236-078-00WHO PQ Public Report1. LabelsPage 12 of 94October 2020, version 6.0
PQDx 0236-078-00WHO PQ Public Report1.1 Outside Box labelPage 13 of 94October 2020, version 6.0
100
PQDx 0236-078-00WHO PQ Public Report1.2 Master lot bar codePage 14 of 94October 2020, version 6.0
HIV-1 Quant Dx AssayMASTER LOT BARCODE SHEETAW-11231 Rev. 003100AMaster Lot No.:123456 PVMaster Lot Exp Date:2021-10-26Master Lot Date of Manufacture:2019-02-25Amplification 021-10-262019-02-232802222222261021Promoter 021111111EPROTCREnzyme ReagentTarget Capture ation Reconstitution Solution5555552021-10-26Enzyme Reconstitution SolutionER6666662021-10-26Promoter Reconstitution SolutionPROR7777772021-10-26Calibrator Coefficient 1280000001122334455Calibrator Coefficient 2280000012233445566Calibrator Coefficient 3280000103344556677Calibrator Coefficient 4280000114455667788
PQDx 0236-078-00WHO PQ Public Report1.3 Calibrator labelOctober 2020, version 6.01.4 Calibrator tray label1.5 Control kit Tray labelPage 15 of 94
PQDx 0236-078-00WHO PQ Public Report1.6 Low positive control labelOctober 2020, version 6.01.7 Negative control label1.8 High positive controlPage 16 of 94
PQDx 0236-078-00WHO PQ Public Report1.9 Amplification reagent1.10 Amplification reconstitution solutionPage 17 of 94October 2020, version 6.0
PQDx 0236-078-00WHO PQ Public Report1.11 Enzyme reagent label1.12 Enzyme reconstitution solution labelPage 18 of 94October 2020, version 6.0
PQDx 0236-078-00WHO PQ Public Re
WHO Prequalification of In Vitro Diagnostics Programme . PUBLIC REPORT . Product: Aptima HIV-1 Quant Dx Assay . WHO Reference number: PQDx . 0236- 078-00. Aptima HIV-1 Quant Dx Assay with product codes -03000 (PRDPRD -03002, PRD-03001), 303014 and PRD-03003, and instrument Panther system with product code 303095,
WHO PREQUALIFICATION PROGRAMME Prequalification is NOT stand alone activity Many other technical work areas support and link to prequalification (medicines, vaccines, diagnostics and medical devices) Outside EMP – Disease oriented departments/programs, IVB Department, Strategic Advisory Group of Experts (SAGE) on
Prequalification of In Vitro Diagnostics Programme - WHO prequalification assessment of products that successfully meet WHO/FIND mRDT product testing programme acceptance criteria through dossier review and site inspection and prequalification for UN procurement. - As of November 2015, WHO has 12 prequalified mRDT products from 4 suppliers: .
The World Health Organization (WHO) Prequalification of In Vitro Diagnostics (IVDs) Programme is coordinated through the department of Essential Medicines and Health Products (EMP). The aim of the WHO Prequalification of IVDs Programme is to promote and facilitate access to safe,
The WHO PQ Programme and its Updated Role after the Outbreaks of Ebola and Zika Virus Robyn Meurant Dept of Essential Medicines and Health Products Prequalification Team –Diagnostics Assessment IMPACT OF REGULATORY REQUIREMENTS: NAT SCREENING. 2 Aim of WHO Prequalification: Diagnostics Assessment To promote and facilitate access
The WHO Programme of Prequalification of Diagnostics and Medical Devices uses the results of the WHO Malaria RDT Product Testing Programme as the laboratory evaluation component of the prequalification process for malaria RDTs. Although not currently a requirement for WHO procurement, manufacturers are encouraged to apply for WHO prequalification.
Consultant Prequalification Application Application Instructions In order to expedite the review time of prequalification applications, all applications must conform to the following requirements: 1. Applications must contain the company name, a current address, email address, phone number, FEIN number, and contact name for correspondence purposes.
(AIA Document A305 – 1986 Contractor’s Qualification Statement) Revised: 10-27-2011 S:\FORMS\OPERATIONS\CONTRACT ADMIN\SUBS & SUPPLIERS PREQUALIFICATION\SUB PACKAGE\2B1S. SUBCONTRACTOR PREQUALIFICATION SUPPLEMENT.DOC When submitting the AIA Document A305
Ulster Archaeological Society at the Divis and Black Mountain site. 5 Illustration 1: Divis Mountain viewed from the south west 1.2 Aims In order to enhance the archaeological record of this site, the aims of this survey were to produce an accurate plan drawing of the monument and carry out a photographic survey. This information was compiled into a report and submitted to the Environment and .
