Foreword
Introduction
1 Scope
2 Normative references
3 Terms and definitions
4 General
5 Characteristics of biological indicators
5.1 General
5.2 Test organism suspension for direct inoculation
of products
5.3 Inoculated carriers
5.4 Self-contained biological indicators
5.5 Other biological indicators
6 Selection of supplier
6.1 General
6.2 Documentation
6.2.1 General
6.2.2 Manufacturer audit
7 Biological indicators in process development
7.1 General
7.2 Overkill approach
7.3 Combined biological indicator and bioburden method
7.4 Bioburden method
8 Biological indicators in sterilization validation
8.1 General
8.2 Placement and handling of biological indicators
8.3 Sterilizer qualification
8.4 Performance qualification
8.5 Review and approval of validation
8.6 Requalification
9 Biological indicators in routine monitoring
9.1 General
9.2 Placement and handling of biological indicators
9.3 Process challenge device (PCD)
10 Results
10.1 General
10.2 Interpretation of results
11 Application of biological indicator standards
11.1 General assessment of biological indicator
performance by the user
11.2 Nominal population of test organism
11.3 Resistance determination
11.3.1 General
11.3.2 Survivor curve method
11.3.3 Fraction-negative method
11.4 z-value determination
11.4.1 General
11.4.2 Graphically plotting the z-value
11.4.3 Mathematically calculating the z-value
11.4.4 Correlation coefficient (r) for the z-value
11.5 F (T, Z) Equivalent sterilization value determination
11.6 Establishing spore-log-reduction (SLR)
11.7 Sterility assurance level (SAL) calculation
11.8 Testing equipment
12 Culture conditions
12.1 General
12.2 Incubation temperature
12.3 Incubation period
12.4 Choice of growth medium
13 Third-party requirements
13.1 General
13.2 Minimum requirements for replicates and total
number of biological indicators
13.3 Test equipment
14 Personnel train
15 Storage and handling
15 Storage and handling
16 Disposal of biological indicators
Annex A (informative) - Microbiological inactivation kinetics
and enumeration techniques
Annex B (informative) - Process challenge devices
B.1 General
B.2 Helices
B.3 Standard test packs
B.4 User's process challenge devices
B.5 Biological test packs
Annex C (informative) - Formulae for fraction negative methods
for D-value calculations
C.1 General
C.2 Materials
C.3 Methods
C.3.1 Holcomb-Spearman-Karber Procedure (HSKP)
C.3.2 Limited Holcomb-Spearman-Karber-Procedure (LHSKP)
C.3.3 The Stumbo-Murphy-Cochran-Procedure (SMCP)
Annex D (informative) - Examples of documentation for biological
indicators prepared by the user
D.1 General
D.1.1 Sources of microorganism
D.1.2 Documentation
D.2 Commercially available suspension
D.3 Suspension from a commercially available strain
D.4 Suspension from in-house isolates
D.5 Inoculated carriers
D.5.1 General
D.5.2 Documentation of fluid carrier materials
D.5.3 Documentation of solid carrier materials
D.5.4 Documentation of inoculated carriers used for
Annex E (informative) - Calculation of z-value
Annex F (informative) - D-value determination by survivor
curve method
F.1 General
F.2 Materials
F.3 Procedure
Bibliography