FINAL DRAFT

International
Standard

ISO/FDIS 7218
ISO/TC 34/SC 9
Microbiology of the food chain —
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General requirements and guidance Secretariat: AFNOR

for microbiological examinations Voting begins on:

2024-01-31
Microbiologie de la chaîne alimentaire — Exigences générales et Voting terminates on:
recommandations pour les examens microbiologiques 2024-03-27

RECIPIENTS OF THIS DRAFT ARE INVITED TO SUBMIT,

WITH THEIR COMMENTS, NOTIFICATION OF ANY
RELEVANT PATENT RIGHTS OF WHICH THEY ARE AWARE
AND TO PROVIDE SUPPOR­TING DOCUMENTATION.
IN ADDITION TO THEIR EVALUATION AS

ISO/CEN PARALLEL PROCESSING BEING ACCEPTABLE FOR INDUSTRIAL, TECHNO­

LOGICAL, COMMERCIAL AND USER PURPOSES, DRAFT
INTERNATIONAL STANDARDS MAY ON OCCASION HAVE
TO BE CONSIDERED IN THE LIGHT OF THEIR POTENTIAL
TO BECOME STAN­DARDS TO WHICH REFERENCE MAY BE
MADE IN NATIONAL REGULATIONS.

Reference number
ISO/FDIS 7218:2024(en) © ISO 2024
 ISO/FDIS 7218:2024(en)
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Contents Page

Foreword................................................................................................................................................................................................................................................... vii
Introduction........................................................................................................................................................................................................................................ viii
1 Scope.............................................................................................................................................................................................................................................. 1
2 Normative references.................................................................................................................................................................................................. 1
3 Terms and definitions................................................................................................................................................................................................. 1
4 Premises.................................................................................................................................................................................................................................... 5
4.1 General.........................................................................................................................................................................................................................5
4.2 Biosafety considerations............................................................................................................................................................................. 5
4.3 Laboratory design............................................................................................................................................................................................. 5
4.4 Laboratory areas................................................................................................................................................................................................ 5
4.4.1 General......................................................................................................................................................................................................5
4.4.2 Areas associated with samples and testing.............................................................................................................6
4.4.3 General areas.......................................................................................................................................................................................6
4.5 Layout and fittings of the premises................................................................................................................................................... 6
4.5.1 Objectives...............................................................................................................................................................................................6
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4.5.2 Fittings......................................................................................................................................................................................................7
4.5.3 Other arrangements for laboratory premises........................................................................................................7
4.5.4 Cleaning and disinfection.........................................................................................................................................................8
5 Personnel.................................................................................................................................................................................................................................. 8
5.1 General.........................................................................................................................................................................................................................8
5.2 Competence............................................................................................................................................................................................................. 8
5.3 Verification of ongoing staff competence..................................................................................................................................... 9
5.4 Hygiene........................................................................................................................................................................................................................ 9
6 Equipment and consumables............................................................................................................................................................................... 9
6.1 General.........................................................................................................................................................................................................................9
6.2 Sterilization and other heating equipment..............................................................................................................................10
6.2.1 General...................................................................................................................................................................................................10
6.2.2 Autoclave..............................................................................................................................................................................................10
6.2.3 Culture media preparator.....................................................................................................................................................11
6.2.4 Steamers, including boiling-water baths................................................................................................................ 12
6.2.5 Sterilizing oven.............................................................................................................................................................................. 12
6.2.6 Microwave oven............................................................................................................................................................................ 13
6.2.7 Hotplate and heating mantle.............................................................................................................................................. 14
6.2.8 Gas burner or wire incinerator......................................................................................................................................... 14
6.3 Temperature controlled equipment and monitoring devices................................................................................. 15
6.3.1 General.................................................................................................................................................................................................. 15
6.3.2 Incubator............................................................................................................................................................................................. 15
6.3.3 Thermostatically controlled bath..................................................................................................................................16
6.3.4 Heating blocks................................................................................................................................................................................. 17
6.3.5 Refrigerators and cold-storage rooms.......................................................................................................................18
6.3.6 Freezer and deep freezer/ultra-low temperature freezer.......................................................................19
6.3.7 Temperature-monitoring devices, including automatic recorders..................................................19
6.3.8 Balances and gravimetric diluters............................................................................................................................... 20
6.4 Defined volume inoculation equipment......................................................................................................................................21
6.4.1 Pipettes and pipettors..............................................................................................................................................................21
6.4.2 Dispensers.......................................................................................................................................................................................... 22
6.4.3 Spiral platers.................................................................................................................................................................................... 23
6.4.4 Serial diluters...................................................................................................................................................................................24
6.5 Protective cabinets........................................................................................................................................................................................24
6.5.1 Description.........................................................................................................................................................................................24
6.5.2 Use............................................................................................................................................................................................................. 25
6.5.3 Cleaning and disinfection..................................................................................................................................................... 25
6.5.4 Maintenance and inspection.............................................................................................................................................. 26

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6.6 Homogenizers, blenders, mixers and shakers...................................................................................................................... 26

6.6.1 Homogenizers and blenders............................................................................................................................................... 26
6.6.2 Vortex mixers...................................................................................................................................................................................27
6.7 Stills, deionizers and reverse-osmosis units......................................................................................................................... 28
6.7.1 Description........................................................................................................................................................................................ 28
6.7.2 Use............................................................................................................................................................................................................. 28
6.7.3 Maintenance..................................................................................................................................................................................... 28
6.7.4 Verification........................................................................................................................................................................................ 28
6.8 Separation and concentration equipment................................................................................................................................ 28
6.8.1 Immunomagnetic separator (IMS)............................................................................................................................... 28
6.8.2 Centrifuge........................................................................................................................................................................................... 29
6.8.3 Filtration systems....................................................................................................................................................................... 29
6.9 Modified atmosphere equipment..................................................................................................................................................... 29
6.9.1 Description........................................................................................................................................................................................ 29
6.9.2 Use............................................................................................................................................................................................................. 29
6.9.3 Maintenance..................................................................................................................................................................................... 30
6.9.4 Verification........................................................................................................................................................................................ 30
6.10 Other equipment............................................................................................................................................................................................. 30
6.10.1 pH meter.............................................................................................................................................................................................. 30
6.10.2 Colony-counting device...........................................................................................................................................................31
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6.10.3 Timers and timing devices...................................................................................................................................................32

6.10.4 Optical microscope......................................................................................................................................................................32
6.10.5 Glass washers, glassware and other laboratory ware................................................................................ 33
6.10.6 Disposable equipment and consumables................................................................................................................ 34
6.10.7 Other equipment and software....................................................................................................................................... 34
7 Sterilization/decontamination and disposal of laboratory materials................................................................. 34
7.1 Sterilization......................................................................................................................................................................................................... 34
7.1.1 General.................................................................................................................................................................................................. 34
7.1.2 Sterilization by dry heat........................................................................................................................................................ 34
7.1.3 Sterilization by moist heat (steam).............................................................................................................................. 34
7.2 Decontamination and disinfection................................................................................................................................................. 34
7.2.1 Decontamination of glassware and materials before use....................................................................... 34
7.2.2 Decontamination of glassware and materials after use........................................................................... 35
7.3 Waste management...................................................................................................................................................................................... 35
7.4 Washing.................................................................................................................................................................................................................. 35
8 Preparation and use of culture media and reagents............................................................................................................... 35
9 Laboratory samples.................................................................................................................................................................................................... 36
9.1 Sampling techniques and sampling plans................................................................................................................................ 36
9.1.1 General.................................................................................................................................................................................................. 36
9.1.2 Sampling.............................................................................................................................................................................................. 36
9.2 Sample transport............................................................................................................................................................................................ 36
9.3 Sample receipt....................................................................................................................................................................................................37
9.4 Sample handling.............................................................................................................................................................................................. 38
9.4.1 General.................................................................................................................................................................................................. 38
9.4.2 Storage before examination............................................................................................................................................... 38
9.4.3 Test portions.................................................................................................................................................................................... 38
9.4.4 Storage of laboratory samples after examination.......................................................................................... 38
9.5 Pre-testing of samples............................................................................................................................................................................... 38
10 Examination........................................................................................................................................................................................................................ 39
10.1 Hygienic precautions during sample preparation and examination................................................................ 39
10.1.1 General.................................................................................................................................................................................................. 39
10.1.2 Basic precautions......................................................................................................................................................................... 39
10.1.3 Sample handling........................................................................................................................................................................... 39
10.1.4 Sample handling tools and implements................................................................................................................... 40
10.1.5 Spillages............................................................................................................................................................................................... 40
10.1.6 Process controls............................................................................................................................................................................ 40
10.1.7 Aerosols................................................................................................................................................................................................ 40

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10.1.8 Molecular methods......................................................................................................................................................................41

10.2 Preparation of initial suspension and dilutions..................................................................................................................41
10.2.1 General...................................................................................................................................................................................................41
10.2.2 Concentration...................................................................................................................................................................................41
11 Enumeration (quantitative) methods...................................................................................................................................................... 41
11.1 General......................................................................................................................................................................................................................41
11.2 Enumeration using a solid medium................................................................................................................................................42
11.2.1 General...................................................................................................................................................................................................42
11.2.2 Pour plate technique..................................................................................................................................................................42
11.2.3 Surface plating techniques...................................................................................................................................................43
11.2.4 Enumeration of yeasts and moulds............................................................................................................................. 44
11.2.5 Incubation...........................................................................................................................................................................................45
11.2.6 Calculation and expression of results obtained with solid culture media................................45
11.2.7 Calculations for enumeration methods.....................................................................................................................47
11.3 Enumeration using liquid media...................................................................................................................................................... 54
11.3.1 Principle............................................................................................................................................................................................... 54
11.3.2 General MPN procedure......................................................................................................................................................... 55
11.3.3 Limitations of MPN..................................................................................................................................................................... 55
11.3.4 Inoculation procedure............................................................................................................................................................. 55
11.3.5 Choice of MPN configuration............................................................................................................................................. 56
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11.3.6 Incubation...........................................................................................................................................................................................57
11.3.7 Interpretation and expression of results................................................................................................................57
11.3.8 Determination of MPN values using MPN calculators.................................................................................57
11.3.9 Rarity categories...........................................................................................................................................................................57
11.4 Estimates of uncertainty of test results.................................................................................................................................... 58
12 Detection (qualitative) methods................................................................................................................................................................... 58
12.1 General..................................................................................................................................................................................................................... 58
12.2 Principle................................................................................................................................................................................................................. 58
13 Confirmation and identification methods.......................................................................................................................................... 59
13.1 General......................................................................................................................................................................................................................59
13.2 Preparation of a pure culture...............................................................................................................................................................59
13.3 Confirmation methods.............................................................................................................................................................................. 60
13.3.1 Latex agglutination test......................................................................................................................................................... 60
13.3.2 Nucleic acid hybridization or molecular amplification methods...................................................... 60
13.3.3 Slide agglutination tests........................................................................................................................................................ 60
13.4 Identification methods............................................................................................................................................................................... 61
13.4.1 Biochemical galleries.................................................................................................................................................................61
13.4.2 DNA sequencing............................................................................................................................................................................. 61
13.4.3 Mass spectrometry..................................................................................................................................................................... 61
14 Selection and characterization of control microorganisms............................................................................................ 62
14.1 General......................................................................................................................................................................................................................62
14.2 Characterization of microorganisms............................................................................................................................................62
14.2.1 General...................................................................................................................................................................................................62
14.2.2 Phenotypic characterization..............................................................................................................................................62
14.2.3 Molecular characterization..................................................................................................................................................62
14.3 Selection of control microorganisms........................................................................................................................................... 63
15 Test report............................................................................................................................................................................................................................ 64
16 Laboratory quality control in microbiology..................................................................................................................................... 64
16.1 General..................................................................................................................................................................................................................... 64
16.2 Internal quality control............................................................................................................................................................................ 65
16.2.1 General.................................................................................................................................................................................................. 65
16.2.2 Process controls............................................................................................................................................................................ 66
16.2.3 Replicate testing............................................................................................................................................................................67
16.2.4 Spiked samples................................................................................................................................................................................67
16.2.5 IQC assessment using control charts..........................................................................................................................67
16.3 External quality assessment.................................................................................................................................................................67

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17 Validation and verification of microbiological methods..................................................................................................... 67

17.1 General......................................................................................................................................................................................................................67
17.2 Performance characteristics................................................................................................................................................................ 68
17.3 Validation.............................................................................................................................................................................................................. 68
17.4 Verification........................................................................................................................................................................................................... 69
Annex A (informative) Properties of disinfectants...................................................................................................................................... 70
Annex B (informative) Confidence intervals for colony count technique............................................................................. 71
Annex C (normative) General confirmation tests.......................................................................................................................................... 74
Bibliography.......................................................................................................................................................................................................................................... 79
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Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out through
ISO technical committees. Each member body interested in a subject for which a technical committee
has been established has the right to be represented on that committee. International organizations,
governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely
with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are described
in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the different types
of ISO documents should be noted. This document was drafted in accordance with the editorial rules of the
ISO/IEC Directives, Part 2 (see www.iso.org/directives).
ISO draws attention to the possibility that the implementation of this document may involve the use of (a)
patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed patent
rights in respect thereof. As of the date of publication of this document, ISO had not received notice of (a)
patent(s) which may be required to implement this document. However, implementers are cautioned that
this may not represent the latest information, which may be obtained from the patent database available at
www.iso.org/patents. ISO shall not be held responsible for identifying any or all such patent rights.
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Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and expressions
related to conformity assessment, as well as information about ISO’s adherence to the World Trade
Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 9,
Microbiology, in collaboration with the European Committee for Standardization (CEN) Technical Committee
CEN/TC 463, Microbiology of the food chain, in accordance with the Agreement on technical cooperation
between ISO and CEN (Vienna Agreement).
This fourth edition cancels and replaces the third edition (ISO 7218:2007), which has been technically
revised. It also incorporates the Amendment ISO 7218:2007/Amd 1:2013.
The main changes are as follows:
— the calculations section has been simplified and two further calculators have been added;
— the equipment section has been reorganized into groups with similar purposes and requirements;
— cross-references have been added to other general microbiology standards such as those for media,
validation and verification, and uncertainty to reduce repetition;
— information on laboratory quality control and characterization of control microorganisms has been
added.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.


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