（英文）行业资料 who -Methods manual for laboratory quality control testing of malaria RDTs

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METHODSMANUALFOR

LABORATORYQUALITYCONTROLTESTINGOFMALARIA

RAPIDDIAGNOSTICTESTS

Manualofstandardoperatingproceduresfor:

Laboratory-basedqualitycontroltestingofmalariarapiddiagnostictestsusingstoreddilutionsofmalariaparasitesand

Preparationofqualitycontrolsamplesfrommalariaparasitefieldcollections

VersionTen

MARCH2023

WHOGlobalMalariaProgramme(GMP),Geneva,Switzerland

Forinternaluseonly

Document:

MalariaRDTQCMethodsManual

Subject:

Introductorynote

RevisionDate:

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INTRODUCTORYNOTE

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MethodsManualforLaboratoryQualityControlTestingofMalariaRDTs

Importantintroductorynote

ThismanualisintendedprimarlyforinternalusebylaboratoriesimplementingWHO-recognizedmalariaRDTlottestingprocedures.

Carefulreferenceshouldbemadetothenotesunder‘ObjectivesandScopeoftheMethodsManual’whenusingthismanual.

ThedesignationsemployedandthepresentationofthematerialinthispublicationdonotimplytheexpressionofanyopinionwhatsoeveronthepartoftheWorldHealthOrganizationconcerningthelegalstatusofanycountry,territory,cityorareaorofitsauthorities,orconcerningthedelimitationofthefrontiersorboundaries.ThementionofspecificcompaniesorofcertainmanufacturersproductsdoesnotimplythattheyareendorsedorrecommendedbytheWorldHealthOrganizationinpreferencetoothersofasimilarnaturethatarenotmentioned.Errorsandomissionsexcepted,thenamesofproprietaryproductsaredistinguishedbyinitialcapitalletters.

TheWorldHealthOrganizationdoesnotwarrantthattheinformationcontainedinthispublicationiscompleteandcorrectandshallnotbeliableforanydamagesincurredasaresultofitsuse.

Document:

MalariaRDTQCMethodsManual

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Acknowledgementsandaddressforcorrespondence

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ACKNOWLEDGEMENTS

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Acknowledgements

ThepreviousversionsofthisproceduremanualweredevelopedbyWHORegionalOfficefortheWesternPacific,WHOGlobalMalariaProgrammeincollaborationwiththeFoundationforInnovativeNewDiagnostics(FIND),andtheUNICEF/UNDP/WorldBank/WHOSpecialProgrammeforResearchandTraininginTropicalDisease(TDR),insupportoftheWHO-FINDmalariaRDTevaluationprogramme.ThisversionwasdevelopedbyWHOGlobalMalariaProgrammeandtheResearchInstituteforTropicalMedicine(RITM).Themanualincludesinputsfromnumeroussources,includingresearchlaboratories,manufacturersanddevelopersofmalariarapiddiagnostictestsandqualityassurancespecialists.

ParticularacknowledgementisduetotheParasitologyDepartmentoftheResearchInstituteforTropicalMedicine(RITM),-DepartmentofHealth,Philippines,;CollegeofPublicHealth,UniversityofPhilippines;;InstitutPasteur,PhnomPenh,Cambodia;theNationalCenterforParasitology,EntomologyandMalariaControl,Cambodia,theMalariaBranch,DivisionofParasiticDiseases,CentersforDiseaseControlandPrevention,Atlanta,USA;theClinicalParasitologyDepartment,HospitalforTropicalDisease,London,UK;theparticipantsforthe2003,2004,2006,2007,2008and2009WHOinformalconsultationsonqualityassuranceofmalariarapiddiagnostictests;RebeccaGuy,BurnetInsititute,Australia;KenLilley,ArmyMalariaInstitute,Australia;KatherineGrady,USCDC;PernilleJorgensen,Berlin,Germany;JenniferLuchavez,RITM,Philippines;andWellingtonOyibo,UniversityofLagos,Nigeria.

Version10includesupdatedproceduresbasedonregularcorrespondencebetweentheWHOprogrammecoordinatorandRITMinthePhilippines,from2017topresent.

Addressforcorrespondence

Mailto:

Malaria_rdt@

KeytoProcedureHistoryrevisions:

AA–AudreyAlbertini,FoundationforInnovativeNewDiagnostics(FIND),Geneva,Switzerland

DB-DavidBell,externalconsultant,Geneva,Switzerland

CS–CharuniSenanayake,Malaria,VectorborneandOtherParasiticDiseases,WHO-RegionalOfficefortheWesternPacific,Philippines

4

CL–ChristianLuna

JB–JohnBarnwell,CentersforDiseaseControlandPrevention,Atlanta,USA

JC–JaneCunningham,WorldHealthOrganization,Geneva,Switzerland

JG–JeffreyGlenn,CentersforDiseaseControlandPrevention,Atlanta,USA

JL–JenniferLuchavez,ResearchInstituteofTropicalMedicine,Muntinlupa,Philippines

KB-KatherineBowers,HospitalforTropicalDiseases,London,UK

KGL-KenLilley,AustralianArmyMalariaInstitute,Brisbane,Australia

NC–NoraChampouillon,FoundationforInnovativeNewDiagnostics(FIND),Geneva,Switzerland

PLC-PeterChiodini,HospitalforTropicalDiseases,London,UK

PJ–PernilleJorgensen,independentconsultant,Berlin,Germany

RG-RebeccaGuy,formerlyMacFarlaneBurnetInstituteforPublicHealthandMedicalResearch,Melbourne,AustraliaRRC–RoxanneRees-Channer,HospitalforTropicalDiseases,London,UK

QC–QinCheng.ArmyMalariaInstitute,Australia

SI–SandraIncardona,FoundationforInnovativeNewDiagnostics(FIND),Geneva,Switzerland

WO–WellingtonOyibo,UniversityofLagos,Lagos,Nigeria

Document:

MalariaRDTQCMethodsManual

Subject:

Tableofcontents

RevisionDate:

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Section:

TABLEOFCONTENTS

Version:

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MethodsManualforLaboratoryQualityControlTestingofMalariaRDTs

TABLEOFCONTENTS

CHAPTER1:INTRODUCTION 10

1.01LISTOFABBREVIATIONS 11

1.02OBJECTIVESANDSCOPEOFTHISMETHODSMANUAL 13

1.03INTRODUCTION 15

1.04MAJORCHANGESFROMVERSION8 18

1.05TERMSOFREFERENCEFORMALARIASPECIMENCOLLECTIONANDRDTLOTQATESTINGSITES 19

1.06LISTOFLABORATORIESINVOLVEDINLABORATORYQCTESTINGOFMALARIARDTS 21

1.07NOTINGANDVARYINGPROCEDURESINTHISMANUAL 23

CHAPTER2:RDTQUALITYCONTROLPROCEDURE 24

SOP2.01OrganizationoftheLotTestinglaboratory 25

SOP2.02Receipt,storageanddispatchofMalariaRDTs 31

SOP2.03RDTQCCommunicationpathway 34

SOP2.04PerforminganRDTwithQCsamplealiquots 38

SOP2.05RDTLotTestingProcedure 41

SOP2.06RDTLaboratoryExternalQualityAssuranceProgramme 56

SOP2.06aRDTLaboratoryEQAProgramme–UseofProficiencyTestingPanels 59

SOP2.06bRDTLaboratoryEQAProgramme–ProductionofProficiencyTestingPanels 62

CHAPTER3:RDTQCSAMPLEPREPARATION 66

PART1:OVERVIEW,REQUIREMENTSANDPREPARATORYACTIVITIES 67

SOP3.01PreparationofQualityControlSamples:Overviewandrequirements 68

SOP3.02PreparationofQualityControlSamples:Preparatoryactivities 75

PART2:PATIENTRECRUITMENTANDBLOODSAMPLECOLLECTIONINTHEFIELD 79

SOP3.03PreparationofQualityControlSamples:Fieldcollectionprocedure 80

SOP3.04TransportandstorageofRDTsinthefield 85

SOP3.05Finger-prickbloodcollectionandpreparationofmalariaRDTsandbloodfilms 88

SOP3.06Venousbloodcollectionandpreparationofbloodfilms 91

SOP3.07Preparationofbloodspotsonfilterpaper,usingfreshfinger-prickorvenousblood 93

PART3:PREPARATIONOFRDTQCSAMPLEDILUTIONSINTHELABORATORY 95

SOP3.08PreparationofQualityControlSamples:DilutionProcedure 96

SOP3.09PreparationofQualityControlSamples:NegativeControlSamples 111

SOP3.10PreparationofQualityControlSamples:Parasite-freebloodfordilution 115

SOP3.11ThawingofFreshFrozenPlasma 122

SOP3.12PerforminganRDTwithfreshlypreparedQCsampledilutions 125

SOP3.13Preparationofbloodspotsonfilterpaper,usingvenousEDTAblood 127

SOP3.14ReversePipettingTechnique 129

SOP3.15LightMicroscopyforRedBloodCellClumping 131

PART4:STORAGEANDTRANSPORTOFRDTQCSAMPLES 134

SOP3.16StorageandTransportofRDTQCsamples 135

SOP3.17PackagingofQualityControlSamplesforTransport 138

SOP3.18DocumentationofQualityControlSamplesforTransport 146

SOP3.19CoordinationofTransportofQualityControlSamples 151

CHAPTER4:MICROSCOPY 154

SOP4.01MalariaMicroscopy:BloodfilmPreparation,StainingandReading 155

SOP4.02MalariaMicroscopy:PreparationandReadingofEarl-Perezslides 166

CHAPTER5:SAMPLECHARACTERIZATION 171

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SOP5.01CellabsPtyHRP2ELISAKitProcedure 172

SOP5.02SDpLDHELISAKitProcedure 178

SOP5.03BiotinylationofMonoclonalAntibodiesforAldolaseELISAProcedure 184

SOP5.04CDCAldolaseELISAformalariaantigeninblood 193

SOP5.05DilutionProtocolforRecombinantpLDH,HRP2,reagentsandbloodsamples(ELISA) 199

SOP5.06ProtocolforRecordingELISAResults 201

SOP5.07ExtractionofGenomicDNAfromWholeBloodUsingQIAampProtocol 204

SOP5.08IdentificationofPlasmodiumSpeciesbyPCRAssay 206

CHAPTER6:GENERALLABORATORYQUALITYASSURANCE 211

SOP6.01LaboratorySafety 212

SOP6.02Training 217

SOP6.03MicroscopeMaintenance 219

SOP6.04MalariaMicroscopyCompetencyAssessment 221

SOP6.05OperationoftheAnalyticalBalance 226

SOP6.06PipetteCalibration 228

SOP6.07IncubatorCalibrationandMaintenance 231

SOP6.08EquipmentTemperatureMonitoring 234

SOP6.09OperationofpHmeter 236

SOP6.10DocumentControl 238

SOP6.11DocumentStorage 241

SOP6.12CorrectiveAction 243

SOP6.13On-siteExternalQualityAssessment(SupervisoryVisits) 245

CHAPTER7:FORMS 248

2.01:RESPONSIBILITIESOFRDT-QCSTAFF 249

2.02:MALARIARDTLOT-TESTREQUESTFORM 251

2.03:RDTFRONTDESKREGISTER(OPTIONAL) 252

2.04:RDTREGISTER 253

2.05:STORAGEANDINTERNALMOVEMENTSOFMALARIARDTS 254

2.06:RDTDISPATCH 255

2.07:RDTQCRESULTSSHEET 256

2.08:RDTQUALITYCONTROLREPORT 259

2.09:ACCESSORYASSESSMENTFORM 264

3.01:PREPARATORYACTIVITIESFORRDTQCSAMPLEPREPARATION 266

3.02:SUPPLIESANDEQUIPMENTSCHECKLIST 268

3.03:STAFFRESPONSIBILITIES 273

3.04A:INFORMATIONSHEETANDCONSENTFORMSFORSTUDYPARTICIPATION 275

3.04B:INFORMATIONSHEETANDCONSENTFORMSFORVIRUSTESTINGANDBLOODSTORAGE 290

3.05:MALARIAPATIENTSCREENINGFORM 308

3.06:PATIENTRECORD 309

3.07:VENEPUNCTURE 311

3.08:PARASITE-FREEBLOODPREPARATION 313

3.09:MALARIAMICROSCOPYRECORD(MICROSCOPIST1,FIRSTREAD) 315

3.10:MALARIAMICROSCOPYRECORD(MICROSCOPIST2,FIRSTREAD) 317

3.11:MALARIAMICROSCOPYRECORD(MICROSCOPIST1,SECONDREAD) 319

3.12:MALARIAMICROSCOPYRECORD(MICROSCOPIST2,SECONDREAD) 321

3.13:PARASITEDENSITY&DILUTIONCALCULATIONS 323

3.14:DILUTIONPREPARATION 325

3.15:RDTRESULTSSHEET 327

3.16:QCSAMPLEPREPARATIONCHECKLISTS 328

3.17:NEGATIVECONTROLSAMPLES 330

3.18:INTERNALMOVEMENTSOFRDTQCSAMPLES 332

3.19:QCSAMPLEREFERRALLOG 333

5.01:ELISADILUTIONFORM 334

5.02:ELISAREPORTINGFORM 335

6.01:PIPETTECALIBRATIONSHEET 338

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6.02:INCUBATORCALIBRATIONSHEET 339

6.03:EQUIPMENTMAINTENANCESHEET 340

6.04:TEMPERATUREMONITORINGFORM 341

6.05:CORRECTIVEACTIONREGISTER 342

Document:

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MethodsManualforLaboratoryQualityControlTestingofMalariaRDTs

LitstofFigures,Tables,andAnnexes

Figures

Figure2-1OverviewofLot-Testing

27

Figure2-2OrganizationandCommunicationPathwayforRDTLot-Testing

36

Figure2-3SummaryofRDTsrequiredforinitialQCforRDTs

43

Figure2-4FlowdiagramofinitialQCtestingofPf-onlyRDTs

48

Figure2-5FlowdiagramofinitialQCtestingofcombinationRDTs(Pf-PanandPf-Pv)

49

Figure2-6AlgorithmforQCtestingofPf-onlyandCombinationRDTs(RDTlot-testingprocedures)

51

Figure2-7AlgorithmforQCtestingofPf-onlyandCombinationRDTs(RDTlaboratoryEQA

programme–usingproficiencytestingpanels)

59

Figure2-8NumberofeachsetofRDTstobeprocuredandpreparedforoneyearofproficiency

testing

62

Figure3-1OverviewoftheRDTQCsamplepreparationprocess

72

Figure3-2Organizationofbloodcollectionfromfield

82

Figure3-3FilterpaperbloodspotsforDNAanalysis

92

Figure3-4SummaryoftheQCsampledilutionprocessinthelaboratory

105

Figure3-5Differentoptionsfortheprocurementandpreparationof“parasite-free

blood”

118

Figure3-6Flowchartforpreparationof“parasite-free”blood

119

Figure3-7FilterpaperbloodspotsforDNAanalysis

127

Figure3-8Packagingofqualitycontrolsamplealiquots

141

Figure3-9Exampleofcorrectlabellingforshipmentofinfectioussubstances(QCsamples,

dangerousgoods,labelclass6)chilledwithdryice(dangerousgoodslabelclass9)

142

Figure3-10Exampleofpackaginglistforattachmenttooutsideofshipment

146

Figure3-11Exampleofacompleteddangerousgoodsformforthetransportationofinfectious

substanceondryice

147

Figure5-1LoadingformatforcoatedpanelsSDHRP2ELISAkitprocedure)

173

Figure5-2Generationoflogarithmicandarithmetictrendlinesfordatainterpretation(SDHRP2

ELISAkitprocedure)

174

Figure5-3Loadingformatforlysisandcoatedplates(SDpLDHELISAkitprocedure)

179

Figure5-4Generationoflogaritmicandarithmetictrendlinesfordatainterpretation(SDpLDHELISA

kitprocedure)

180

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Figure5-5Loadingformatforlysisandcoatedplates(CDCAldolaseELISAformalariaantigenin

blood)

194

Figure5-6Generationoflogarithmicandarithmetictrendlinesfordatainterpretation(CDCAldolase

ELISAformalariaantigeninblood)

196

Tables

Table2-1Numberoflot-RDTsthatmustbepositiveinInitialQCtestingfortheRDTlottopass

50

Table3-1Summaryofaliquotsandtests(PreparationofQCsamples)

106

Table3-2Summaryofforms(PreparationofQCsamples)

106

Table5-1RecombinantPfHRP2(xng/ml)*variesdependingonaliquotsbeingused(SDHRP2

ELISAkitprocedure)

172

Table5-2HazardouschemicalsusedintheCellabsmalariaantigenHRP2ELISA(SDHRP2ELISA

kitprocedure)

175

Table5-3RecombinantpLDH(xng/ml)*variesdependingonaliquotsbeingused(SDpLDHELISA

kitprocedure)

178

Table5-4HazardouschemicalsusedintheSDmalariaantigenpLDHELISA(SDpLDHELISAkit

procedure)

181

Table5-5RecombinantPvAldolase(xng/ml)*variesdependingonaliquotsbeingused(SDpLDH

ELISAkitprocedure)

193

Table5-6HazardouschemicalsusedintheCDCaldolaseELISA

196

Table5-7Checklistofinformationtoberecordedinthehardcoverlaboratorybook(Protocolfor

recordingELISAresults)

200

Table5-8Nest1PCRMasterMix

206

Table5-9Nest1ThermalProfile

207

Table5-10Nest2PCRMasterMix

207

Table5-11Nest2ThermalProfile

208

Table5-12GenusandSpecies-SpecificPrimerPairsforNest1andNest2PCRReactions

208

Annexes

Annex1:RDTIntensityRatingChartsforStabilityAssessment

363

Annex2:GuideFortheInterpretationofObservationsNotedDuringLotTestingofMalaria

364

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MethodsManualforLaboratory

QualityControlTestingofMalaria

RDTs

Chapter1:INTRODUCTION

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Listofabbreviations

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1.01ListofAbbreviations

TermExplanation

Ab

Antibody

Ag

Antigen

AAMI

ArmyMalariaInstitute(Queensland,Australia)

CDC

CentersforDiseaseControlandPrevention(Atlanta,UnitedStatesofAmerica)

CIDEIM

CentroInternacionaldeEntrenamientoyInvestigacionesMédicas(Cali,Colombia)

CNM

NationalCenterforParasitology,EntomologyandMalariaControl

(PhnomPenh,Cambodia)

DMR

ExperimentalMedicineResearchDivision

(DepartmentofMedicalResearch,Yangon,Myanmar)

DNA

DesoxyribonucleicAcid

EDTA

EthylenediamineTetra-aceticAcid

EHTH

EthiopianHealth&NutritionResearchInstitute(AddisAbeba,Ethiopia)

ELISA

Enzyme-linkedImmunosorbentAssay

EQA

ExternalQualityAssurance

EQALAT

ExternalQualityAssuranceLaboratoryAssessmentTool

EQAP

ExternalQualityAssurancePanel

EQC

ExternalQualityControl

FIND

FoundationforInnovativeNewDiagnostics

GE

GeneralExternalQualtiyAssuranceIndicator

HIV

HumanImmunodeficiencyVirus

HRP2

Histidine-richProtein2

HTD

HospitalforTropicalDiseases

(London,UnitedKingdomofGreatBritainandIreland)

ID

Identificationnumber

IHRDC

IfakaraHealthResearchandDevelopmentCenter(Bagamoyo,Tanzania)

IMT

InstitutodeMedicinaTropical(UniversidadPeruanaCayetanoHeredia,Lima,Peru)

IATA

InternationalAirTransportAssociation

IPB

InstitutPasteurdeBangui(Bangui,CentralAfricanRepublic)

IPC

InstitutPasteurduCambodge(PhnomPenh,Cambodia)

IPM

InstitutPasteurdeMadagascar(Antananarivo,Madagascar)

IQC

InternalQualityControl

KEMRI

KenyaMedicalResearchInstitute(Kisumu,Kenya)

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MVP

Malaria,vector-borneandotherparasiticdiseases

Non-Pf

NonPlasmodiumfalciparumspecies

(Plasmodiumvivax,Plasmodiummalariae,Plasmodiumovale)

Pan

Plasmodium

PCR

PolymeraseChainReaction

Pf

Plasmodiumfalciparum

pLDH

PlasmodiumLactateDehydrogenase

Pm

Plasmodiummalariae

Po

Plasmodiumovale

Pv

Plasmodiumvivax

p/µL

Parasitespermicrolitre

QA

QualityAssurance

QC

QualityControl

RDT

RapidDiagnosticTest.Forthepurposesofthismanual,thisreferstoimmunochromatographiclateralflowdevicesforthedetectionofmalariaparasiteantigens

RITM

ResearchInstituteforTropicalMedicine(Manila,Philippines)

SD

StandardDiagnostics(Seoul,SouthKorea)

SOP

StandardOperatingProcedure

TDR

UNICEF/UNDP/WorldBank/WHOSpecialProgrammeforResearchandTraininginTropicalDiseases

UL

UniversityofLagos(Lagos,Nigeria)

WHO

WorldHealthOrganization

WPRO

WesternPacificRegionalOffice

‘Lot-testing

OfficerdesignatedbyWHOtocoordinateoveralllot-testingprogramme,or

Coordinator’

officerauthorizedtoactonbehalfofthatperson

Project

OfficerdesignatedbyWHOtomanagethelot-testingprogramme,orofficer

Manager

authorizedtoactonbehalfofthatperson

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Objectivesandscope,layout

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