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Estimation of measurement uncertainty in food microbiology: The ISO

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Article  in  Accreditation and Quality Assurance · April 2006

DOI: 10.1007/s00769-005-0085-5

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Accred Qual Assur (2006) 17: 94–100
DOI 10.1007/s00769-005-0085-5 LEGISLATION AND NORMS

Bertrand Lombard
Estimation of measurement uncertainty
in food microbiology: The ISO approach

Received: 13 May 2005
Accepted: 19 December 2005
Published online: 14 February 2006


C Springer-Verlag 2006
Presented at AOAC Europe/Eurachem
Symposium March 2005, Brussels, Belgium
B. Lombard (
)
AFSSA-LERQAP (Agence Française
de Sécurité Sanitaire de Aliments–
Laboratoire d’Etudes et de Recherches
sur la Qualité des Aliments et sur les
Procédés Agro-alimentaires),
23 avenue du Général De Gaulle,
94 706 Maisons-Alfort, France
e-mail: b.lombard@afssa.fr
Tel.: +33-1-49-77-26-96
Fax: +33-1-43-68-97-62
Abstract Given the current interest
in measurement uncertainty (MU) in
food microbiology, in particular for
laboratory accreditation purposes,
and the need to have harmonized
reference documents specifically in
that area at the international level,
ISO is conducting works to meet this
need. An ISO Technical Specification
(ISO/TS 19036) is being prepared on
MU estimation for quantitative
determinations. A global approach
has been chosen, based on the
reproducibility standard deviation of
the final result of the measurement
process. Three possibilities are
envisaged for the estimation of the
reproducibility standard deviation, in
a decreasing order of preference: The
intra-laboratory standard deviation,
the inter-laboratory standard
deviation derived from method
validation, and the inter-laboratory
standard deviation derived from
proficiency testing.The uncertainty of
qualitative determinations is still
under investigation, and will be
covered by a separate ISO
publication.
Keywords Food . Microbiology .
Measurement uncertainty . ISO

Introduction the finalization of international standardization works that

Microbiological laboratories, like the overall analytical
community, are more and more addressing the question
of measurement uncertainty (MU) attached to the results
they produce. The main reason for this trend is certainly
to be found in the frame of laboratory accreditation, where
the Standard EN ISO 17025 [1], in its clause 5.4.6, re-
quires laboratories to estimate MU associated to their re-
sults, and there is no reason that this general requirement
would not apply to microbiology. The same standard also
states that if rigorous/statistically valid calculation of MU is
not possible, the MU components are to be identified, and
to be reasonably estimated. The requirements regarding
MU for the accreditation of microbiological laboratories
have been clarified at European level by the European co-
operation for Accreditation (EA) in the Guide EA-04/10
[2]. In France, the body in charge of laboratory accred-
itation, COFRAC (Comité Français d’Accréditation) has
recognized that these requirements should apply to food
microbiology; it requests that laboratories address the ques-
tion of MU, identify and control the main sources of un-
certainty, but it has delayed the need to estimate MU up to
are presented in this article. In addition to the accreditation
requirements, the particular significance of microbiologi-
cal analysis may be emphasized: It detects or quantifies in
many cases a direct hazard for consumers’ heath, making
it all the more critical to assess the level of confidence that
can be put in the outcome of the analysis. The variability
associated with many reference methods used to determine
colony counts in food microbiology is large, typically in the
0.1–1.0 log10 cfu/g range (Table 7.6 in [3]). In particular
for the interpretation of the results, the need to estimate and
control this large variability is thus reinforced. Finally, most
analyses for food pathogens are currently performed on a
presence/absence basis. The way to express the uncertainty
of qualitative determinations needs to be investigated, no
solution being currently available.
These reasons have led ISO/TC 34/SC 9 “Food
products—Microbiology” to address the question of MU
in food microbiology at its Bangkok meeting in December
2002. ISO/TC 34/SC 9 is in charge of standardization in the
field of food microbiology, preparing the so-called “hori-
zontal” ISO Standards, which are applicable to the analysis
of all foods and feeds, including the environment of food
production and food handling. At the 2002 meeting, it has

been decided to prepare at first the ISO guidelines on MU
estimation in quantitative microbiology. MU for qualitative
determinations would be dealt with in a second step, since
this second topic would require more investigation before
an approach could be agreed upon.
Measurement uncertainty in quantitative microbiology
Presentation
The member countries and international organisations in
liaison of ISO/TC 34/SC 9 have agreed on the need to
develop guidelines specific to food microbiology, even if
a large set of reference documents was already available
to address and estimate MU of quantitative determinations
from a more general point of view, such as the basic refer-
ence document on MU published by ISO, GUM [4], as well
as ISO/TS 21748 [5] or the EURACHEM/CITAC Guide
[6]. ISO/TC 34/SC 9 considered that it was necessary to
address the specificities of microbiology, the analyte being
a living microorganism and some main reagents used in
enumeration techniques (such as culture media, antibod-
ies for confirmation) being of biological nature, resulting
in the large variability of the results recalled earlier in the
introduction. In addition, it can be noted that the existing
ISO/CEN Standards describing reference methods in food
microbiology provide at best, in a limited number of cases,
values of precision data derived from inter-laboratory trials
(see later), but no limits to the values of the major sources
of MU, which could have enabled laboratories to avoid
to estimate MU themselves, according to EN ISO 17025
(Note 2 in clause 5.4.6.2).
Moreover, the need to have a reference document harmo-
nized at the international level was recognized by ISO/TC
34/SC 9. The variety of existing documents introducing dif-
ferent approaches was regarded as a reason to produce an
harmonized reference document in food microbiology. The
intention was to avoid that each country, accreditation body
or even each laboratory would adopt its own approach, that
could lead to marked inconsistency in the approaches to
MU and to the difficulty of comparing MU values from
different laboratories for the same determination (differ-
ences could arise from differences in the approach to MU,
and not from differences in the analytical method chosen
nor in the proficiency of the laboratories).
At its Bangkok meeting, ISO/TC 34/SC 9 decided to es-
tablish an ISO Technical Specification, ISO/TS 19036 [7],
and not an International Standard. It should be noted that
a Technical Specification cannot be regarded as having the
same status as an International Standard, that it cannot have
the same degree of reference and that competing Techni-
cal Specifications on the same subject are permitted [8].
One reason for this choice is that such a status enables a
quicker publication for a topic that was considered as one of
the first priorities of the committee. In addition, the status
of Technical Specification requires a 3-year revision. This
relatively short life time was found appropriate for a topic
that has only recently raised interest in the community of
95
food microbiologists and where the approach selected has
not yet been widely used. Thus, it justifies the choice of
a status of reference document lower than an International
Standard, being still “tentative” for a limited period of time
and needing a review at short-term. Finally, this status is
meant to encourage the users of the Technical Specification
to report their experience on the implementation of the ap-
proach described. After the 3-year period, ISO/TS 19036
would then be reviewed by ISO/TC 34/SC 9 in the light of
the experience gained. After improvements and sufficient
support by users, it is intended to become an International
Standard. In particular, certain aspects not dealt with in the
first edition would need to be addressed, such as MU at low
levels of contamination and MU of Most Probable Number
techniques. In addition, a choice between the three options
left open to estimate the reproducibility standard deviation
may have to be made (see later).
The principle of the approach was easily agreed upon by
ISO/TC 34/SC 9 at its 2002 meeting, with a broad consen-
sus of countries and observing international organizations,
based on a first series of trials performed earlier in the year
that had aimed at testing the feasibility of the approach
proposed.
ISO/TS 19036 applies not only to colony-count tech-
niques, the reference methods in quantitative food micro-
biology, but also to other quantitative techniques regarded
as alternative methods, which can be of instrumental nature
(such as impedancemetry, ATP-metry, flow cytometry,. . .)
and whose signal is converted into number of colonies, the
unit of the reference methods.
The approach selected
Two main approaches can be distinguished to estimate MU
for any type of measurement. The first one consists in esti-
mating the individual sources of variability (variances) that
contribute to the uncertainty in the measurement process,
and MU is derived using formal principles of uncertainty
propagation by combination of variances. This “bottom
up” or “step-by-step” approach is a widely adopted stan-
dard approach, described in the GUM as well as in several
other publications, particularly in a more practical way by
the EURACHEM/CITAC Guide for analytical chemistry.
It has been applied also to quantitative microbiology by the
Finnish Centre for Metrology and Accreditation (MIKES)
[9]. This approach can be rather easily implemented in
water microbiology, where the matrix is relatively simple
and homogeneous. However, this approach has not been
recommended by ISO/TC 34/SC 9 for food microbiology,
in accordance with other reference documents and authors
(such as ISO/TS 21748, Horwitz [10]). The following rea-
sons can be given to justify this choice:
1. The bottom up approach, to be implemented routinely,
requires the use of standard or assumed variances for
the individual steps that align the combined estimate to
the basic method rather than the analytical result.
96
2. ISO/TC 34/SC 9 considered that such a process might
underestimate the true MU value in the case of the micro-
biological analysis of food where it is difficult to build
a really comprehensive model of the measurement pro-
cess. The measurement process of colony-count tech-
niques is complex and includes several steps, the im-
portant ones being: Sub-sampling of the test portion,
preparation of the initial suspension and of the serial
decimal dilutions, plating on isolation agars, incubation,
counting of colonies, confirmation step in some cases. A
significant source of variation may thus be overlooked.
3. It appears difficult to quantify accurately the contribu-
tion of each individual step of the analytical process
in food microbiology, where (a) the analyte is a liv-
ing microorganism, whose physiological state can be
largely variable, (b) the analytical target includes dif-
ferent strains, or different species, or different gen-
era, and (c) the enumeration techniques use reagents
of biological nature (such as culture media, antibod-
ies for serological confirmation) that may largely vary
in an unpredictable manner. In other words, it is diffi-
cult to build a metrologically rigorous and statistically
valid estimation of MU for the microbiological anal-
yses of foods, as it has been recognized by the EA
Guide 04/10.
4. Some error factors can be unpredictable, such as drop-
ping the thermometer or missing a decimal point, and
are not subject to a predictive statistical description that
is needed in the decomposition approach (Horwitz [10]).
5. A purely modelling approach would not take account of
the variability due to difference in sample types and to
the sub-sampling of the test portion. A restricted view
to MU may exclude the sub-sampling of the test por-
tion from the estimation of MU that would be limited
to the further analytical steps. A larger view includes in
MU all steps performed by the laboratory, and in food
microbiology, the heterogeneous distribution of the mi-
croorganisms in most food types (solid ones) can be the
source of a major part of the total uncertainty, at the
sub-sampling step.
6. Given the complexity of the measurement process in
food microbiology, the step-by-step approach is rather
heavy to implement in this analytical field.
ISO/TC 34/SC 9 has chosen the second approach, based
on the reproducibility standard deviation (sR ) of the final
result of the entire measurement process, here the number
of colonies per gram or millilitre of sample. This experi-
mental approach makes no attempt to set generic estimates
of uncertainty for specific test methods and rightly aligns
the MU estimate with a specific analysis, or set of anal-
yses. It is likely that most of the potential variability fac-
tors that are likely to be encountered in practice, including
the sub-sampling of the test portion and the unpredictable
tricks, will be introduced (Horwitz, [10]). This “global” or
“top down” approach has been endorsed by different refer-
ence structures, including Codex Alimentarius (the Codex
Committee on Methods of Analysis and Sampling [11]),
and ISO/TC 69/SC 6 “Application of statistical methods—
Measurement methods and results,” which has prepared
ISO/TS 21748, a publication that creates a bridge between
GUM and the ISO 5725 series dedicated to the estimation
of trueness and precision of analytical methods, and that
justifies the consistency of the global approach with GUM.
The global approach can be considered as a special case of
GUM’s type A experimental estimation. Finally, the Inter-
national Laboratory Accreditation Cooperation (ILAC) in
its general guide on MU in the context of ISO/IEC 17025
[12], encourages the use of existing experimental data from
quality control charts or inter-laboratory trials, which are
at the basis of the global approach.
The main steps
In May 2003, the first draft of ISO/TS 19036 was prepared
on the basis of the first series of trials of 2002 and the
decisions taken in Bangkok, the author acting as Project
Leader for ISO/TC 34/SC 9.
From December 2003 to March 2004, a second series of
ISO trials has been conducted. Their purpose was to quan-
tify the MU component linked to (i) the sub-sampling of the
test portion, and (ii) the preparation of the initial suspension
(see details later). The burden was shared by 79 laboratories
from 10 countries, a level of participation not yet encoun-
tered in trials launched by ISO/TC 34/SC 9, denoting the
interest of the analytical community for that topic!
Based on the comments received from on member coun-
tries and organizations in liaison, and the outcome of the tri-
als, a Project Group recommended in a meeting during 22–
23 March 2004, modifications to the first draft in order to
prepare the final draft of ISO/TS 19036, and these were
endorsed by ISO/TC 34/SC 9 at its meeting in Parma,
April 2004.
The Technical Specification ISO/TS 19036
The final draft of ISO/TS 19036 has been submitted to
the official voting period until 11 May, and it will soon be
published after being modified according to the comments
received during the vote.
Right from the title of ISO/TS 19036, the intention to
define a “guide” or guidelines is expressed. It means that
ISO/TC 34/SC 9 was conscious that other publications or
reference documents are available on the same topic, and
that the approach of ISO/TS 19036 is not the unique nor the
mandatory approach to be followed in this field is unique,
this approach being nevertheless the recommended one in
food microbiology. The choice of a Technical Specifica-
tion status had a similar objective (see earlier). Moreover,
the guidelines mention in the introduction the step-by-step
approach as well as its application to microbiology.
As in any ISO reference document, one of the first sec-
tions provides definitions, including MU itself. They are all
extracted from GUM.
The Technical Specification then describes the global
approach, which can be conveniently illustrated by a

Fig. 1 Diagram of the main Excluded (out of
sources of uncertainty in food the analytical
microbiology, and the process)
“black-box” approach to
measurement uncertainty
Sampling
Laboratory
sample
Matrix
Low levels excluded
“black box” diagram as in Fig. 1. This diagram mentions
the main uncertainty sources that are covered, partly
covered or not covered by the approach chosen, such as
sampling, excluded from the measurement process senso
stricto. Regarding the bias, part of it can be estimated
in the experimental conditions of the intra-laboratory
reproducibility and to a larger extent in the conditions of
the inter-laboratory reproducibility (see later). However,
the empirical nature of microbial counts, where the analyte
is defined by the method used, justifies that in the general
case the total bias cannot be included in MU estimation,
the true value being unknown.
ISO/TS 19036 insists that the global approach is to be
implemented in a laboratory under quality control, as to
ensure the validity of MU estimation in the routine ana-
lytical activity, and not only at a given time. It means that
the critical factors associated with the method or the lab-
oratory that are likely to affect the measurement result,
should be identified and demonstrated to be under control.
A re-assessment of MU estimation is required following
changes of any of these factors. In other words, the global
approach does not preclude that the laboratory identifies
the main components of uncertainty, in agreement with the
requirements of ISO/IEC 17025, but the quantification of
each component is not required.
The following principles are then defined:
(i) Expanded MU (U)=2 sR , the usual value of 2 being
chosen for the coverage factor (95% confidence level);
(ii) sR is to be estimated per microorganism (or consistent
group of), per matrix type (or consistent group of), for
a given physiological state of the organism and for a
given method that the laboratory uses for producing in
routine its results. The term “consistent” means that
the group would give equivalent MU values.
(iii) Three options are recommended for estimating sR
• Intra-laboratory sR ;
• Inter-laboratory sR derived from method validation;
• Inter-laboratory sR derived from proficiency testing.
97
Serial Counting/
Sub-sampling/ dilutions/ confirmation
Primary dilution plating of colonies
Black box Result
Residual Operator/ Equipment/ Bias
random time culture
errors media/
reagents Total bias not covered
(empirical nature of
microbial counts)
ISO/TS 19036 does not deal with MU estimation at low
levels (<10 colonies counted on a plate) due to a lack of
a simple agreed approach, but it is expected that the next
edition of these guidelines would cover it.
Intra-laboratory sR
The intra-laboratory sR is a particular case of “intermediate
precision” as defined in the Standard ISO 5725-3 [13]; the
reproducibility conditions are varied within a laboratory:
Analysis of the same sample in conditions differing as much
as possible within the same laboratory (see details later).
This is the preferred option, since it provides an MU value
that is linked to the laboratory, thus nearer to the basic MU
definition which relates MU even to one individual test
result.
An experimental protocol is described in ISO/TS 19036,
and is represented in Fig. 2. From the same food sample,
two test portions are taken by two different operators (the
“operator” may be a team of operators—technicians, each
of whom performing a given part of the procedure). From
each test portion, one initial suspension is prepared, and
analysed once, as in routine testing. Conditions A and B
should be as different as possible and should ideally include
as many variations as may be encountered from one day
to another within the laboratory, in terms of technicians,
batches of culture media and reagents, vortex mixer, pH
meter, incubators, time of analysis, etc. If the contamina-
tion of the food sample is demonstrated to be sufficiently
stable (which is rarely the case in food microbiology), the
conditions A and B should relate also to different days of
analysis. The purpose is to cover the extremes of method-
ological variability likely to occur in a given laboratory
in order to derive a maximum estimate of MU that would
reflect the range of individual results likely to be obtained
in the same laboratory. The laboratory can then safely as-
sociate this MU value with its results without the risk of
underestimating MU.
98
Food sample
1st operator (conditions A) 2nd operator (conditions B)
Initial suspension Artificial contamination
(if needed)
Analysis
Different conditions
Fig. 2 Experimental protocol for the intra-laboratory standard de-
viation of reproducibility
This protocol is to be repeated for at least 10 samples for
a given combination (target microorganism, matrix type);
a consistent group of each of them can also be considered.
The number of matrix types to be tested depends on the
diversity of the matrices analysed in routine by the labo-
ratory. The selected matrices should be representative, in
terms of MU values, of the matrix types analysed by the
laboratory and should also be relevant to the microorgan-
isms for which the test is to be done. This selection is
certainly the most challenging aspect in this approach, and
an inappropriate choice would lead to an unrealistic MU
estimation. Moreover, it makes the protocol less practicable
when the laboratories analyse a large variety of matrices,
each requiring a separate MU assessment (such as in the
case of official food control laboratories).
This protocol incorporates the effect of the sub-sampling
of the test portion in the evaluation of the total uncertainty.
Since it is well known in food microbiology that the nat-
ural contamination of certain food products can be highly
heterogeneous, this MU component may have a major con-
tribution to the total MU and cannot be avoided, especially
when a judgement on conformity of the sample analysed
against limits (such as microbiological criteria) is to be
made.
ISO/TS 19036 then provides the different calculations
up to MU, not detailed here. The data are initially log-
transformed, to stabilize the reproducibility variance over
the contamination levels, given that low levels are not con-
sidered here.
Inter-laboratory sR derived from method validation
If the method used routinely by the laboratory has been
submitted to an inter-laboratory study for validation, the
laboratory may use the reproducibility standard deviation
of the method for deriving its own MU under the following
conditions:
1. The laboratory bias is compatible with that expected on
the basis of the repeatability and reproducibility esti-
mates derived from the inter-laboratory study;
2. The precision attained by the measurements within the
laboratory is compatible with that expected from re-
peatability and reproducibility estimates derived from
the inter-laboratory study;
3. The inter-laboratory study has correctly covered all the
Initial suspension sources of uncertainty (especially sample preparation
and homogenisation).
Analysis
These pre-requisites are justified by the fact that the stan-
dard deviation derived from an inter-laboratory study is
linked to the method, and not to a given laboratory which
is to report MU attached to its results. The procedure to
check that these conditions are met, and how to form a
combined uncertainty estimate with the possible additional
factors not covered by the inter-laboratory study, are not
described in detail in ISO/TS 19036 which refers for these
aspects to ISO/TS 21748.
This approach has the advantages to use precision data
already generated and to enable a laboratory that has taken
part in an inter-laboratory study to assess its laboratory
bias, part of the bias component of MU. This aspect is not
detailed in ISO/TS 19036.
However, in food microbiology, there are several limita-
tions to this approach, justifying its consideration only as a
second option:
– In addition to the need to check the pre-requisites listed
before, only a limited number (5) of reproducibility pa-
rameters have been derived from inter-laboratory studies
for standardized reference methods in food microbiol-
ogy.
– It may be difficult to generalise values obtained in an
inter-laboratory study to the routine samples analysed
by the laboratory. Precision values of an inter-laboratory
study will be obtained under a limited and precisely de-
fined combination of matrices, strains and physiological
state of microorganism, contaminations level, etc. and
for a given background microflora (if present).
– Given the homogeneity requirements for the samples
used for inter-laboratory studies, the need to send to the
laboratories homogenised and stabilised samples induces
a reduction of the “natural” variation in sample con-
tamination that may be found in practice, which causes
underestimation of the uncertainty.
Inter-laboratory sR derived from proficiency testing
If the laboratory has taken part in an inter-laboratory pro-
ficiency testing (PT), it may use the standard deviation of
reproducibility from this PT to deduce its measurement
uncertainty, under the following conditions:
– During the inter-laboratory study, the laboratory used the
same method that it uses in routine analyses;
– The samples that were used in the study are comparable,
in terms of matrix and contamination level, to the ones
analysed routinely;
– The laboratories that participated in the study did not use
different empirical methods, or a sufficient number of

participants used the same method, as to allow a correct
estimation of the reproducibility standard deviation.
This option presents the two same advantages than
the former option. However, most of the limitations
encountered for the former option apply here too, except
the limited availability of data, given the current large
number of PT schemes organized in food microbiology.
Expression of measurement uncertainty in test reports
ISO/TS 19036 proposes several possibilities to express
MU, including the following:
– y±2sR , where y=log10 x is the test result, sR expressed
in log units,
– after back transformation, x[10 y−2sR , 10 y+2sR ].
Results of trials
An informative annex of ISO/TS 19036 provides the results
of the trials organized in 2003/2004 for ISO/TC 34/SC
9, which aimed at estimating the uncertainty component
linked to the sub-sampling of the test portion and to the
preparation of the initial suspension. The protocol and the
results of these trials are detailed into a report prepared by
the trials’ organizers [14] and are intended to be published
in a separate paper.
Each of the 79 participating laboratories elected to enu-
merate one or more target microorganisms in one or more
matrix types naturally contaminated with the target mi-
croorganisms of interest. The participants followed a com-
mon experimental protocol laid down for the trials, corre-
sponding to the protocol of ISO/TS 19036 for the intra-
laboratory sR (see Fig. 2) except that for each condition
A or B, two duplicates were tested under repeatability
conditions. The protocol was also to be repeated for 10
samples of the same combination (matrix type, target mi-
croorganism). Ninety-six combinations (laboratory, target
microorganism, matrix type) meeting the acceptability cri-
teria defined for the trials were used to calculate, for each
combination, the intra-laboratory sR , as well as, with the
theory of the variance components, the three standard devi-
ations linked to (i) sub-sampling of test portion/preparation
of initial suspension, (ii) the intra-laboratory conditions
(operator/time/reagents/equipment) as defined by ISO/TS
19036 in the experimental protocol for the intra-laboratory
sR , and (iii) the residual errors (random errors under re-
peatability conditions).
A classification of matrices related to physical criteria
was tested: (i) liquids and powders (e.g. milk, coconut milk,
dried milk); (ii) well-mixed solids (e.g. minced meat, me-
chanically separated meat, sausage meat, crushed meat,
whipped cream, dairy ice cream, soya cream); (iii) small
(or very small) solids (e.g. dehydrated parsley/mushrooms,
grated carrots/celeriac, salad, shrimps, cereals, feeding
stuffs, chopped hazel nuts); and (iv) other solids (e.g. non-
minced meat, cheeses, pastry). Using a non-parametric
99
Table 1 Values of standard deviation linked to sub-sampling of
test portion/preparation of initial suspension (sIS ), in log10 cfu/g, per
food category (2003–2004 trials)
Food Minimum Median Maximum Mean (sIS )
category (sIS ) (sIS ) (sIS )
I 0.02 0.08 0.17 0.08
II 0.01 0.09 0.17 0.09
III 0.08 0.17 0.41 0.19
IV 0.05 0.20 0.74 0.25
Note. I: liquids and powders; II: well-mixed solids; III: small (or very
small) solids; and IV: other solids
ANOVA (Kruskall–Wallis by ranks), the factor category
was shown to have a significant impact (p=5.10−7 ) on
the values of the MU component value linked to sub-
sampling/initial suspension.
Table 1 gives the values of the standard deviation linked
to sub-sampling of test portion/preparation of initial sus-
pension for each of the food categories defined above.
For the first two categories, the matrix was responsible for
about 0.1 log units of the overall standard deviation, regard-
less of the laboratory or the microorganism. For the last two
categories, it was not possible to assess the order of mag-
nitude of the effect independent of the microorganism and
the laboratory. On the basis of these trials, ISO/TC 34/SC
9 concluded that it was not possible to decompose total
MU between the component linked to sub-sampling/initial
suspension (a typical value per type of matrix) and the
component linked to the following analytical steps (to be
determined by each laboratory).
Analytical uncertainty in qualitative microbiology
Strictly speaking, a qualitative test is not a measurement,
but it is still possible to characterize the uncertainty in the
result of a qualitative determination, the term “analytical
uncertainty” may be more appropriate in that context. The
need to characterize the uncertainty of detection methods
(presence/absence tests) is particularly important in food
microbiology, as highlighted before, since most of the con-
trols on food-borne pathogens are currently conducted on
a qualitative basis.
This section is not expanded, since preliminary works
are still under way, conducted by Working Group 2 “Statis-
tics” of ISO/TC 34/SC 9. For detection methods, it is en-
visaged to follow the proposal from one of the group’s
members, Hitchins (non-published), deriving the analyti-
cal uncertainty from the 50% limit of detection (LOD50% ).
This is the value of the contamination level where a posi-
tive result is obtained in 50% of the cases. This point cor-
responds to the inflexion point of the sigmoid curve (dose,
response) well-investigated in toxicological studies, and for
which several models have been developed. In particular,
WG 2 of ISO/TC 34/SC 9 is currently investigating the
model which would be best suited to microbiological de-
tection methods, on the basis of experimental data already
generated.
100

Conclusion for other analytical fields where qualitative determinations

The global approach retained at ISO level to estimate MU
for quantitative determinations appears to be pragmatic,
adapted to the complexity of the analytical process in food
analysis, especially in food microbiology. It is in agreement
with some international reference documents and rules.
It is foreseen that ISO will soon agree on a way to express
the uncertainty of presence/absence determinations, that
would be of interest not only for food microbiology, but also
are performed.
It is hoped that the publication of ISO/TS 19036 will
encourage the generalization of MU estimation in food
microbiology, thus leading the analytical field of food mi-
crobiology from an empirical state to a more “scientific”
one!
Acknowledgements The author wishes to thank Dr Marie Cornu
and Professor Basil Jarvis for their help in improving the text of the
publication.

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