ForewordIntroductionScopeNormative referencesTerms and definitionsGeneralCharacteristics of biological indicatorsGeneralTest organism suspension for direct inoculation of productsInoculated carriersSelf-contained biological indicatorsOther biological indicatorsSelection of supplierGeneralDocumentationGeneralManufacturer auditBiological indicators in process developmentGeneralOverkill approachCombined biological indicator and bioburden methodBioburden methodBiological indicators in sterilization validationGeneralPlacement and handling of biological indicatorsSterilizer qualificationPerformance qualificationReview and approval of validationRequalificationBiological indicators in routine monitoringGeneralPlacement and handling of biological indicatorsProcess challenge device (PCD)ResultsGeneralInterpretation of resultsApplication of biological indicator standardsGeneral assessment of biological indicator performance by the userNominal population of test organismResistance determinationGeneralSurvivor curve methodFraction-negative methodSurvival-kill response characteristicsz value determinationGeneralGraphically plotting the z valueMathematically calculating the z valueCorrelation coefficient, r, for the z valueF equivalent sterilization value determinationEstablishing spore-log-reduction (SLR)Sterility assurance level (SAL) calculationTest equipmentCulture conditionsGeneralIncubation temperatureIncubation periodChoice of growth mediumThird-party requirementsGeneralMinimum requirements for replicates and total number of biological indicatorsTest equipmentPersonnel trainingStorage and handlingDisposal of biological indicatorsMicrobiological inactivation kinetics and enumeration techniques (informative)Process challenge devices (informative)GeneralHelicesStandard test packsUser's process challenge devicesBiological test packsFormulae for fraction negative methods for D value calculations(adapted from ) (informative)GeneralMaterialsMethodsHolcomb-Spearman-Karber Procedure (HSKP)IntroductionCalculations using the HSKPExample calculation of the Holcomb-Spearman-Karber Procedure (HSKP)Limited Holcomb-Spearman-Karber Procedure (LHSKP)Calculations using the LHSKPExample calculations of the Limited Holcomb-Spearman-Karber Procedure (LHSKP)Stumbo-Murphy-Cochran Procedure (SMCP)IntroductionCalculations using the SMCPExample calculations of the Stumbo-Murphy-Cochran Procedure (SMCP)Examples of documentation for biological indicators prepared by the user (informative)GeneralSources of microorganismsDocumentationCommercially available suspensionSuspension from a commercially available strainSuspension from in-house isolatesInoculated carriersGeneralDocumentation of fluid carrier materialsDocumentation of solid carrier materialsDocumentation of inoculated carriers used for D value determinationsCalculation of z value(adapted from ) (informative)D value determination by survivor curve method(adapted from ) (informative)GeneralMaterialsProcedureSurvival-kill response characteristics (adapted from ) (informative)GeneralMaterialsMethodBibliography