A24 – CALA CHECKLIST FOR

MICROBIOLOGY
Revision 5.1
March 29, 2023

Laboratory Name:

Appendix Name: ________

Appendix Number: ________

Assessor: ________

Date:
 A24 – CALA CHECKLIST FOR MICROBIOLOGY

SELECTION AND VERIFICATION OF METHODS (7.2.1)

Clause Requirements Observation prior the Observation on-site
visit
7.2.1.1 The laboratory uses appropriate methods and supporting procedures,
including but not limited to testing, sampling, verification and
validation, and estimation of measurement uncertainty procedures.
7.2.1.2 An approved, documented method is authorized (8.3), up-to-date,
available to personnel, and authorized.
Any supporting work instructions (e.g., glassware cleaning, sample
disposal, report of adverse results, etc.) are documented, authorized,
up-to-date and readily available to the analyst.
Any equipment manuals are readily available to the analyst.
7.2.1.3 The method is based on the latest valid edition of a published
reference method (where appropriate and possible). As well, the
method is adequately documented and supplemented with additional
details to ensure consistent application.
Steps to look for include, but are not limited to:
• Sampling;
• Details on reagent preparation, storage and shelf life;
• Procedure for media preparation, including labeling, storage,
quality control and safety procedures for handling of media;
• Colony counting and reporting criteria; appropriate reporting of
non- detects, taking dilution factors and sample volumes into
consideration;
• Equipment and supplies;
• Maintenance of stock cultures.
(see CALA document P07, section 7.2.1.3)
7.2.1.4 Methods are published either in international, regional or national
standards, or by reputable technical organizations, or in relevant
scientific texts or journals, or as specified by the manufacturer of the
equipment.
7.2.1.5 Where there has been no modification from the reference method, the
laboratory has verified that it can properly perform the method prior to
analysis of customer samples.

Revision 5.1 Page 1 of 14

 A24 – CALA CHECKLIST FOR MICROBIOLOGY

For water testing: Requirements will vary on whether the method is

qualitative or quantitative. Refer to CALA Document P07, Appendix 1,
Clause 7.2.1.5.
For food-testing: refer to CALA document P07, Appendix 1, Clause
7.2.1.5.
7.2.1.6 Method development is planned and assigned to competent personnel.
Periodic review is carried out confirm the needs of customer are still
being fulfilled.
Any modifications are approved and authorized.
7.2.1.7 Deviations from methods are documented, technically justified,
authorized and accepted by the customer. See CALA document P07.
VALIDATION OF METHODS (7.2.2)
Clause Requirements Observation prior the visit Observation on-site

7.2.2.1 In-house developed methods, published methods without validation

data, commercial test kits without validation data and
standard/reference methods used outside their intended scope or that
have been modified are appropriately validated. The level and rigour of
validation will depend on the nature/extent of the modification(s).
For Food-Testing requirements, see CALA document P07, section
7.2.2.1.
7.2.2.2 When changes are made to a validated method, influence of such
changes is determined and if found to affect the original validation, a
new method validation is performed.
7.2.2.3 Performance characteristics of validated methods are relevant to the
customers’ needs and consistent with specific requirements.
7.2.2.4 Validation records are retained, including: the validation procedure;
specification of the requirement; determination of the performance
characteristics; results obtained; and, statement on the validity,
detailing its fitness for the intended use.
EVALUATION OF MEASUREMENT UNCERTAINTY (7.6)
Clause Requirements Observation prior the Observation on-site
visit
7.6.1 For all tests, factors are identified which affect measurement
uncertainty.

Revision 5.1 Page 2 of 14

 A24 – CALA CHECKLIST FOR MICROBIOLOGY

7.6.2 For internal calibrations on balances, thermometers and pipettes, there

is a documented procedure that includes the estimation of
measurement uncertainty and staff with the appropriate training.
7.6.3 For quantitative tests, the laboratory has calculated measurement
uncertainty (see CALA Policy P19).
For qualitative tests, the laboratory demonstrates knowledge of the
factors affecting measurement uncertainty and has an understanding
of the false positives/false negatives, where applicable.
SAMPLING (7.3)
Clause Requirements Observation prior the Observation on-site
visit
7.3.1 A sampling plan and method is established and available at the site of
sampling, addresses the factors to be controlled, and is based on
statistical methods, whenever reasonable.
7.3.2 The sampling method describes the selection of samples or sites, the
sampling plan, and the preparation and treatment of samples to yield a
representative sample for testing.
7.3.3 Records with respect to sampling are retained and include, where
relevant:
• Reference to the sampling method used
• Date and time of sampling
• Data to identify and describe the sample (e.g. number,
amount, name)
• Identification of the personnel and equipment
• Environmental or transport conditions
• Diagrams or other equivalent means to identify locations,
when appropriate
• Deviations, additions to or exclusions

Revision 5.1 Page 3 of 14

 A24 – CALA CHECKLIST FOR MICROBIOLOGY

HANDLING OF TEST OR CALIBRATION ITEMS (7.4)

Clause Requirements Observation prior the visit Observation on-site

7.4.1 There are documented procedures for transportation, receipt, handling,

protection, storage, retention and disposal or return of samples,
including all provisions necessary to protect the integrity and interests
of the laboratory and customers. These procedures include, but are
not limited to:
• use of sterile approved sample containers;
• for water testing only: collection of chlorinated samples in
sterilized containers pretreated with sodium thiosulphate
(where appropriate);
• instruction to ensure that holding time and sample
transport / storage meets method / regulatory
requirements;
• For Food testing: temperature at reception recorded (e.g.
10°C for some types of testing)
Precautions are taken to avoid deterioration, contamination, loss or
damage.
Handling instructions are followed.
7.4.2 Samples have a unique, unambiguous identification that is retained
during the testing and reporting process.
7.4.3 Deviations from specified conditions are recorded (e.g. samples does
not fit, sample leaking).
Consultations with customers are recorded.
A disclaimer in the report is recorded when the customer is asking for a
deviation.
7.4.4 Storage conditions are maintained, monitored and recorded.
FACILITIES AND ENVIRONMENTAL CONDITIONS (6.3)
Clause Requirements Observation prior the visit Observation on-site

6.3.1 Environmental conditions are suitable for the laboratory activities.

Conditions do not adversely affect the validity of results.
6.3.2 Requirements for environmental conditions are documented (e.g.,
incubation temperatures).
6.3.3 The laboratory monitors, controls, and records environmental
Revision 5.1 Page 4 of 14
 A24 – CALA CHECKLIST FOR MICROBIOLOGY

conditions with relevant specifications, methods or procedure or

where they influence the validity of the results (e.g., monitoring of
incubators, fridges, etc…).
6.3.4 Measures to control facilities are implemented, monitored and
periodically reviewed including but not limited to:
• Access to laboratory activities
• Prevention of contamination, interference or adverse
influences
• Effective separation of incompatible activities (for example,
this is especially important for PCR methods).
For food-testing only:
Monitoring of the Air quality and records are retained.
Monitoring of the Surface quality (swabs) and records are retained.
Monitoring is performed for the appropriate bacteria.
6.3.5 If activities are performed outside the facilities, ensure the
requirements of the standard are met.
EQUIPMENT (6.4)
Clause Requirements Observation prior the visit Observation on-site

6.4.1 All equipment, reagents, media, reference materials, consumables

and auxiliary apparatus (including software) required for the test
procedure is available, appropriately monitored, and functioning
properly. The laboratory has sufficient quantities of equipment and
supplies to carry out the volume of work. In microbiology, these
items can include, but are not limited to:
• sterile rinse buffer / distilled water;
• disinfectants available and routinely used for cleaning bench
areas;
• reference standards (e.g., reference weights, pH standards,
etc.);
• stock cultures (e.g. ATCC organisms).
6.4.2 When using equipment outside the laboratory’s permanent control,
ensure that the requirements of ISO/IEC 17025 are met.
6.4.3 The procedure and/or supporting procedures include instructions to
ensure that equipment, media, reagents, test organisms, etc. are
handled, transported, stored, and used in a manner to ensure proper
Revision 5.1 Page 5 of 14
 A24 – CALA CHECKLIST FOR MICROBIOLOGY

functioning and to prevent contamination or deterioration. This

includes but is not limited to:
a) When washing and re-using glassware that can impact the
recovery of organisms, the laboratory requires a procedure to
test for residual detergent. See CALA document P07,
Appendix 1 for more direction or guidance.
b) b) Procedures for planned maintenance are in place for
equipment that requires regular servicing or checks.
6.4.4 Equipment was verified prior to being put into use or if it was
returned to service. Supplies that need to be checked prior to use
include but are not limited to:
a) A minimum of 1 sample container from each lot of new,
certified containers is checked for sterility or for influence on
parameters.
b) Purchased organisms have certificates with the organism
name, and the laboratory confirms the identify of these
organisms by either using an acceptable identification method
(e.g. API, Biolog, Vitek, etc.) or demonstrating key reactions on
selective medium.
c) Organisms isolated from the environment are properly
characterized, and key reactions are demonstrated on
selective medium as required.
d) For Water testing only
e) For MF methods, hydrophobicity of filters is tested (e.g.,
“charcoal” test or confluent growth or other method) and
inhibitory effects of filters is tested by comparison of
recoveries on a membrane filter and a spread plate/pour
plate (or equivalent).
f) If producing water in house and it is used to dilute samples, or
make media or reagents, check conductivity daily or as-used
and have procedures in place, including sterility testing, to
verify that the water is not having a negative impact on the
conduct of the test (see CALA document P07, Appendix 1).
g) g) If purchasing distilled water, and it is used to dilute samples
or to make media or reagents, have procedures in place,
including sterility testing, to verify that the water is not having
a negative impact on the conduct of the test. A certificate shall
Revision 5.1 Page 6 of 14
 A24 – CALA CHECKLIST FOR MICROBIOLOGY

be obtained from the supplier prior to use (see CALA

document P07, Appendix 1).
6.4.5 Equipment for measurement is capable of achieving measurement
accuracy and/or measurement uncertainty (e.g., incubators, working
thermometers, etc.).
6.4.6 Equipment is calibrated, if the measurement accuracy or
measurement uncertainty affects the test result or to establish
traceability (e.g., thermometers, balances).
6.4.7 A calibration programme is established, reviewed and adjusted to
maintain confidence in the status of calibrations.
6.4.8 Equipment that requires calibration (i.e. semi-automated pipettes,
balances and thermometers that are critical to the test result) is
labeled to indicate the status, including the date last calibrated and
expiry criteria or date when due. Note: labeling is not required for
equipment verified daily or as-used.
All information required to properly identify organisms appear on
their containers (i.e., name or number of organisms, and date
subcultured) including working cultures and those stored at lower
temperatures (e.g. in refrigerators / freezers, where appropriate).
6.4.9 Equipment that is out-of-service is isolated or marked as being out of
service.
6.4.10 Intermediate checks on equipment are carried out when necessary.
6.4.11 Reference values and correction factors are updated and
implemented, as appropriate (e.g., a correction factor on a working
thermometer).
6.4.12 Practicable measures are taken to prevent unintended adjustments
of equipment that would invalidate results.
6.4.13 Records are retained for equipment, as applicable. Records generally
include: identity of the equipment; the manufacturer’s name, serial
number, or other unique identification; evidence that the equipment
conforms with specified requirements; current location; calibration
dates, results of calibrations, adjustments, acceptance criteria, and
the due date of the next calibration; maintenance plan and
maintenance carried out to date; and details of any unplanned work
or repair on the equipment.
Records shall be maintained of reference materials, results,
Revision 5.1 Page 7 of 14
 A24 – CALA CHECKLIST FOR MICROBIOLOGY

acceptance criteria, relevant dates and the period of validity of

reference materials.

METROLOGICAL TRACEABILITY (6.5)

Clause Requirements Observation prior the Observation on-site
visit
6.5.1 The laboratory has established and maintained metrological
traceability of measurement results (unbroken chain) and
measurement uncertainty of each step (see CALA Policy A61-01).
MEDIA HANDLING AND QUALITY CONTROL
Clause Requirements Observation prior the visit Observation on-site

6.4.8 Media and / or reagents are appropriately labeled with material,

concentration or purity (as required), date prepared and /or expiry
date.
6.4.3 Media/ reagents are stored under proper conditions, and storage
times are met.

Revision 5.1 Page 8 of 14

 A24 – CALA CHECKLIST FOR MICROBIOLOGY

7.5.1 Records are kept for all media/ reagents prepared or received and
include:
• date of receipt;
• date opened / first use;
• date of preparation, and expiry date (if required);
• lot number, as required;
• shelf-life / expiry date of product;
• storage conditions;
• performance specifications.
For purchased / prepared media / reagents: records are retained of
manufacturer’s QC and other information, including but not limited
to positive and negative control(s), sterility and final pH
For in-house prepared media: QC results and sufficient information
to enable the test to be repeated under conditions as close as
possible to the original.
QC records are retained for each batch of in-house prepared or
purchased media.
NOTE: for guidance on defining a batch, refer to the Interpretation
in CALA document P07, Appendix 1. The type and nature of QC
testing will depend on the method.
7.7.1.c Media QC includes a positive control culture (traceable to ATCC or
equivalent).
For Water testing only -
QC must be performed using the same technique as used for
routine analysis. For example, the positive control must be applied
using MF for MF methods.
As well, a comparison of positive control cultures on selective and
non-selective media is done and there is a comparison of recovery
rates of the positive control culture (see P07, Appendix 1 for further
information; not applicable to tube MPN methods).
For other MPN methods (such as quantitray or similar), the lab must
demonstrate satisfactory recovery of target organisms (e.g., using
selective media tray vs non-selective media or old vs new lot
comparisons, or use of quantifiable cultures)
7.7.1.c Media QC includes a sterility check.

Revision 5.1 Page 9 of 14

 A24 – CALA CHECKLIST FOR MICROBIOLOGY

7.7.1.c Media QC includes a negative control culture (traceable to ATCC or

equivalent).
For Water testing only - The negative culture can be streaked on the
media for MF methods.
7.7.1.c For Water testing only -
For quantitative methods using only non-selective media: compare
recovery rates on an old/previous batch of media to a new batch,
using the same technique to do test (e.g., MF or spread plate or
pour plate).
ENSURING THE VALIDITY OF RESULTS (7.7)
Clause Requirements Observation prior the visit Observation on-site

7.7.1 A procedure exists to monitor the validity of the results.

Resulting data is recorded in a way as to detect trends, and where
practicable, statistical techniques are applied to review results (e.g.,
control charts).

7.7.1.f Method/analytical QC includes duplicates to monitor within-run

precision for quantitative methods (where appropriate). For further
guidance, refer to P07, Appendix 1.

7.7.1.g Method/analytical QC includes confirmation of isolates as necessary

(applicable only to membrane filtration methods).

7.7.1.h There are procedures in place to ensure there is no carryover

between membrane filtrations (e.g., UV boxes, hot water).

7.7.1.j Method/analytical QC includes monthly inter-technician comparison

readings to monitor precision and monthly parallel analyses on at
least one positive sample to monitor inter-technician method
precision for quantitative tests.
Note: Inter-technician comparison readings are generally applicable
to any method where a result depends on the judgment of an
analyst (e.g., color reaction).

Revision 5.1 Page 10 of 14

 A24 – CALA CHECKLIST FOR MICROBIOLOGY

7.7.2 Laboratory meets CALA PT Policy for Accreditation (see CALA Policy
P02-03).
7.7.3 QC results are analyzed and used to control and, if applicable,
improve the laboratory’s activities.
If QC results are outside pre-defined criteria, appropriate action is
taken.
If PT did not meet acceptance criteria, confirm that the laboratory
has records of corrective action.
REPORTING OF RESULTS (7.8)
Clause Requirements Observation prior the visit Observation on-sit

7.8.1.1 The results are reviewed and authorized prior to release. For
observations related to reporting, cite the appropriate clause in the
main checklist (A02 – CALA Rating Guide).
7.8.1.2 The results are provided accurately, clearly, unambiguously and
objectively. For observations related to reporting, cite the
appropriate clause in the main checklist (A02 – CALA Rating Guide).

Revision 5.1 Page 11 of 14

 A24 – CALA CHECKLIST FOR MICROBIOLOGY

TECHNICAL RECORDS (7.5)

Clause Requirements Observation prior the visit Observation on-site

7.5.1 Technical records are complete with respect to the specific tests,
are recorded at the time they are made and that there is sufficient
information to establish an audit trail.
Specific records to review include but are not limited to: analyst
worksheet or notebook; records of nonconformities; stock culture
maintenance logs; records of reference standards (e.g. reference
weights, pH standards, etc.) and reference materials certificates
(e.g. ATCC strains).
For guidance on the type and nature of records that may be
needed, please refer to CALA document P07, Appendix 1, Clause
7.5.1- Technical Records.
7.5.2 Amendments to technical records are tracked to previous or
original observations. Both shall be retained including the date of
alteration, an indication of the altered aspects and personnel
responsible.
CONTROL OF DATA AND INFORMATION MANAGEMENT (7.11.2)
Clause Requirements Observation prior the visit Observation on-site

7.11.2 Software (e.g. LIMS) is validated for functionality before

introduction and any changes to software (e.g. LIMS) are
authorized, documented and validated before implementation.
For any observations, cite the appropriate clause in the main
checklist (A02 – CALA Rating Guide).

Revision 5.1 Page 12 of 14

 A24 – CALA CHECKLIST FOR MICROBIOLOGY

OTHER RESOURCES - PERSONNEL (6.2) AND EXTERNALLY PROVIDED PRODUCTS AND SERVICES (6.6)
Clause Requirements Observation prior the visit Observation on-site

6.2 Personnel are competent and the laboratory has authorized

personnel to perform specific lab activities including but not
limited to:
• Development, modification, verification and validations
of methods
• Analysis of results including statement of conformity and
opinions and interpretations
• Reporting, reviewing and authorization of results
For any observations related to personnel, cite the appropriate
clause in the main checklist (A02 – CALA Rating Guide).
6.6 For any observations related to purchasing equipment, supplies or
services, cite the appropriate clause in the main checklist (A02 -
CALA Rating Guide).

Revision 5.1 Page 13 of 14

 A24 – CALA CHECKLIST FOR MICROBIOLOGY

Area Appendix # Appendix # Appendix # Appendix # Appendix # Appendix #

6.2 – Personnel

6.3 – Environmental
Conditions
6.4 – Equipment

6.5 – Traceability

6.6 – Externally provided

products and services
7.2.1 – Method Selection
& Verification
7.2.2- Validation

7.3 - Sampling

7.4 – Sample Handling

7.5 – Technical Records

7.6 - MU

7.7 - Trending

7.7 – Media Prep & QC

7.7 – Method QC (e.g.,

dups, intertech etc.)
7.7 – Confirmation

7.7 – PT

Revision 5.1 Page 14 of 14