ASTM E1326-15
(Guide)Standard Guide for Evaluating Non-culture Microbiological Tests Used for Enumerating Bacteria
Standard Guide for Evaluating Non-culture Microbiological Tests Used for Enumerating Bacteria
SIGNIFICANCE AND USE
5.1 This guide should be used by producers and potential producers of non-culture tests to determine the accuracy, selectivity, specificity, and reproducibility of the tests, as defined in Practice E691. Results of such studies should identify the limitations and indicate the utility or applicability of the non-culture test, or both, for use on different types of samples.
5.2 Non-culture test users and potential users should employ this guide to evaluate results of the non-culture test as compared to their present methods. Practices D5245 and D5465 should be reviewed in regards to the microbiological methods employed. If culture methods have not been used for monitoring the systems, then guidelines are included for obtaining microbiological expertise.
5.3 Utilization of a non-culture test can reduce the time required to determine the microbiological status of the system and detect microbe that are not detected by culture testing. Consequently, non-culture tests can contribute to the improvement in the overall operating efficiency of microbial contamination condition monitoring and diagnostic efforts, and microbicide performance evaluations.
5.4 Detecting microbial contamination levels that exceed predetermined upper control limits indicates the need for an addition of an antimicrobial agent or other corrective maintenance action. By accurately determining this in a shorter time period than is possible than by culture methods, treatment with antimicrobial agents may circumvent more serious problems than if the treatment were postponed until culture results were available. If the antimicrobial treatment program relied on an inaccurate non-culture test, then unnecessary loss of product and problems associated with inappropriate selection or improper dosing with antimicrobial agents would exist.
5.5 Since many methods based on entirely different chemical and microbiological principles are considered, it is not possible to establish a unique design and recomm...
SCOPE
1.1 The purpose of this guide is to assist users and producers of non-culture tests in determining the applicability of the test for processing different types of samples and evaluating the accuracy of the results. Culture test procedures such as the Heterotrophic (Standard) Plate Count, the Most Probable Number (MPN) method and the Spread Plate Count are widely cited and accepted for the enumeration of microorganisms. However, these methods have their limitations, such as performance time and degree of accuracy. Moreover any given culture test method typically recovers only a fraction of the total viable microbes present in a sample. It is these limitations that have recently led to the marketing of a variety of non-culture procedures, test kits and instruments.
1.2 Culture test methods estimate microbial population densities based on the ability of mircoorganisms in a sample to proliferate in or on a specified growth medium, under specified growth conditions. Non-culture test methods attempt to provide the same or complimentary information through the measurement of a different parameter. This guide is designed to assist investigators in assessing the accuracy and precision of non-culture methods intended for the determination of microbial population densities or activities.
1.3 It is recognized that the Heterotrophic Plate Count (HPC) does not recover all microorganisms present in a product or a system (1, 2).2 When this problem occurs during the characterization of a microbiological population, alternative standard enumeration procedures may be necessary, as in the case of sulfate-reducing bacteria. At other times, chemical methods that measure the rates of appearance of metabolic derivatives, the utilization of contaminated product components or genetic profile of the microbial population might be indicated. In evaluating non-culture tests, it is possible that the use of these alternative standard procedures m...
General Information
Relations
Buy Standard
Standards Content (Sample)
NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation: E1326 − 15
StandardGuide for
Evaluating Non-culture Microbiological Tests Used for
1
Enumerating Bacteria
This standard is issued under the fixed designation E1326; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope means available for establishing correlation. In such cases, this
guide can serve as a reference for those considerations.
1.1 Thepurposeofthisguideistoassistusersandproducers
of non-culture tests in determining the applicability of the test 1.4 Because there are so many types of tests that could be
for processing different types of samples and evaluating the considered non-culture based, it is impossible to recommend a
accuracy of the results. Culture test procedures such as the specific test protocol with statistical analyses for evaluating the
Heterotrophic (Standard) Plate Count, the Most Probable tests. Instead, this guide should assist in determining what
Number (MPN) method and the Spread Plate Count are widely types of tests should be considered to verify the utility and
cited and accepted for the enumeration of microorganisms. identify the limitations of the nonconventional test.
However, these methods have their limitations, such as perfor-
1.5 The values stated in SI units are to be regarded as
mance time and degree of accuracy. Moreover any given
standard. No other units of measurement are included in this
culture test method typically recovers only a fraction of the
standard.
total viable microbes present in a sample. It is these limitations
that have recently led to the marketing of a variety of 2. Referenced Documents
non-culture procedures, test kits and instruments.
3
2.1 ASTM Standards:
1.2 Culture test methods estimate microbial population D1129 Terminology Relating to Water
densities based on the ability of mircoorganisms in a sample to
D4012 Test Method forAdenosineTriphosphate (ATP) Con-
proliferate in or on a specified growth medium, under specified tent of Microorganisms in Water
growth conditions. Non-culture test methods attempt to pro-
D5245 Practice for Cleaning Laboratory Glassware,
vide the same or complimentary information through the Plasticware, and Equipment Used in Microbiological
measurementofadifferentparameter.Thisguideisdesignedto
Analyses
assist investigators in assessing the accuracy and precision of
D5465 Practice for Determining Microbial Colony Counts
non-culture methods intended for the determination of micro-
from Waters Analyzed by Plating Methods
bial population densities or activities.
E177 Practice for Use of the Terms Precision and Bias in
ASTM Test Methods
1.3 It is recognized that the Heterotrophic Plate Count
E691 Practice for Conducting an Interlaboratory Study to
(HPC) does not recover all microorganisms present in a
2
Determine the Precision of a Test Method
product or a system (1, 2). When this problem occurs during
E1601 Practice for Conducting an Interlaboratory Study to
the characterization of a microbiological population, alterna-
Evaluate the Performance of an Analytical Method
tive standard enumeration procedures may be necessary, as in
E2756 Terminology Relating to Antimicrobial and Antiviral
the case of sulfate-reducing bacteria. At other times, chemical
Agents
methods that measure the rates of appearance of metabolic
derivatives, the utilization of contaminated product compo-
3. Terminology
nents or genetic profile of the microbial population might be
3.1 Defintions:
indicated. In evaluating non-culture tests, it is possible that the
3.1.1 For definitions of terms used in this guide refer to
use of these alternative standard procedures might be the only
Terminologies D1129, E2756, and E177.
3.2 Abbreviations:
1
This guide is under the jurisdiction of ASTM Committee E35 on Pesticides,
3.2.1 HPC—Heterotrophic Plate Count
Antimicrobials, and Alternative Control Agents and is the direct responsibility of
Subcommittee E35.15 on Antimicrobial Agents.
Current edition approved May 1, 2015. Published June 2015. Originally
3
approved in 1990. Last previous edition approved in 2013 as E1326 – 13. DOI: For referenced ASTM standards, visit the ASTM website, www.astm.org, or
10.1520/E1326-15. contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
2
The boldface numbers in parentheses refer to the list of references at the end of Standards volume information, refer to the standard’s Document Summary page on
this guide. the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 -----------------
...
This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: E1326 − 13 E1326 − 15
Standard Guide for
Evaluating NonconventionalNon-culture Microbiological
1
Tests Used for Enumerating Bacteria
This standard is issued under the fixed designation E1326; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope
1.1 The purpose of this guide is to assist users and producers of nonconventionalnon-culture tests in determining the
applicability of the test for processing different types of samples and evaluating the accuracy of the results. Conventional Culture
test procedures such as the Heterotrophic (Standard) Plate Count, the Most Probable Number (MPN) method and the Spread Plate
Count are widely cited and accepted for the enumeration of microorganisms. However, these methods have their limitations, such
as performance time and degree of accuracy. Moreover any given culture test method typically recovers only a fraction of the total
viable microbes present in a sample. It is these limitations that have recently led to the marketing of a variety of
non-conventionalnon-culture procedures, test kits and instruments.
1.2 A conventional test is one that is widely accepted and published as a standard microbiological method or related procedure.
A new, nonconventional test method will Culture test methods estimate microbial population densities based on the ability of
mircoorganisms in a sample to proliferate in or on a specified growth medium, under specified growth conditions. Non-culture test
methods attempt to provide the same or complimentary information through the measurement of a different parameter. This guide
is designed to assist investigators in assessing the accuracy and precision of nonconventionalnon-culture methods intended for the
determination of microbial population densities or activities.
1.3 It is recognized that the Heterotrophic Plate Count (HPC) does not recover all microorganisms present in a product or a
2
system (1, 2). When this problem occurs during the characterization of a microbiological population, alternative standard
enumeration procedures may be necessary, as in the case of sulfate-reducing bacteria. At other times, chemical methods that
measure the rates of appearance of metabolic derivatives or derivatives, the utilization of contaminated product components or
genetic profile of the microbial population might be indicated. In evaluating nonconventional tests, non-culture tests, it is possible
that the use of these alternative standard procedures maymight be the only means available for establishing correlation. In such
cases, this guide can serve as a reference for those considerations.
1.4 SinceBecause there are so many types of tests that could be considered nonconventional, non-culture based, it is impossible
to recommend a specific test protocol with statistical analyses for evaluating the tests. Instead, this guide should assist in
determining what types of tests should be considered to verify the utility and identify the limitations of the nonconventional test.
1.5 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
2. Referenced Documents
3
2.1 ASTM Standards:
D1129 Terminology Relating to Water
4
D3870 Practice for Establishing Performance Characteristics for Colony Counting Methods in Microbiology (Withdrawn 2000)
D4012 Test Method for Adenosine Triphosphate (ATP) Content of Microorganisms in Water
D5245 Practice for Cleaning Laboratory Glassware, Plasticware, and Equipment Used in Microbiological Analyses
D5465 Practice for Determining Microbial Colony Counts from Waters Analyzed by Plating Methods
D7687E177 Test Method Practice for Measurement of Cellular Adenosine Triphosphate in Fuel, Fuel/Water Mixtures, and
Fuel-Associated Water with Sample Concentration by FiltrationUse of the Terms Precision and Bias in ASTM Test Methods
1
This guide is under the jurisdiction of ASTM Committee E35 on Pesticides, Antimicrobials, and Alternative Control Agents and is the direct responsibility of
Subcommittee E35.15 on Antimicrobial Agents.
Current edition approved Oct. 1, 2013May 1, 2015. Published October 2013June 2015. Originally approved in 1990. Last previous edition approved in 20082013 as
E1326 – 08.E1326 – 13. DOI: 10.1520/E1326-13.10.1520/E1326-15.
2
The boldface numbers
...
Questions, Comments and Discussion
Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.