NELAC V1M5 Microbiology 2012-0717

• Audit Title

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  external audit

1239) : Introduction

• 1240) V1M5 1.1: This Standard applies to laboratories undertaking microbiological analysis of environmental samples. Microbiological testing refers to and includes the detection, isolation, enumeration, or identification of microorganisms and/or their metabolites, or determination of the presence or absence of growth in materials and media. The evaluation of laboratories for this discipline is in conjunction with a quality system as specified in general requirements module. Adherence to quality systems requirements will ensure that all quality control procedures specified in this module are being followed.

1241) V1M5 1.2: Scope

• 1242) V1M5 1.2: The essential quality control procedures applicable to microbiological analysis are included in this module. Additional quality control requirements that are either specified by method, regulation or project shall be met by laboratories.

1243) V1M5 1.3: Terms and Definitions

• 1244) V1M5 1.3: The relevant definitions from TNI, Volume 1, Module 2, Section 3.0 apply. Definitions related to this document, which are used differently or do not exist in the above references are defined below.

• 1245) V1M5 1.3.1: Additional Terms and Definitions

• 1246) V1M5 1.3.1: Reserved

• 1247) V1M5 1.3.2: Exclusions and Exceptions

• 1248) V1M5 1.3.2: Reserved

1249) V1M5 1.4: Method Selection

• 1250) V1M5 1.4: A reference method is a method issued by an organization generally recognized as competent to do so. (When ISO refers to a standard method, that term is equivalent to reference method).

• 1251) V1M5 1.4: When a laboratory is required to analyze a parameter by a specified method due to a regulatory requirement, the parameter/method combination is recognized as a reference method.

• 1252) V1M5 1.4: If there is not a regulatory requirement for the parameter/method combination, the parameter/method combination is recognized as a reference method if it can be analyzed by another similar reference method of the same matrix and technology.

• 1253) V1M5 1.4: When it is necessary to use methods not covered by reference methods, these shall be subject to agreement with the client and shall include a clear specification of the client's requirements and the purpose of the environmental test.

  Yes

  No

  N/A

• 1254) V1M5 1.4: The method developed shall have been validated appropriately before use.

  Yes

  No

  N/A

1255) V1M5 1.5: Method Validation

• 1256) V1M5 1.5: Validation is the confirmation by examination and the objective evidence that the particular requirements for a specific intended use are fulfilled.

• 1257) V1M5 1.5: The laboratory shall validate non-reference methods, laboratory-designed/developed methods, reference methods used outside their published scope, and amplifications and modifications of reference methods to confirm that the methods are fit for the intended use.

  Yes

  No

  N/A

• 1258) V1M5 1.5: The validation shall be as extensive as is necessary to meet the needs of the given application or field of application.

  Yes

  No

  N/A

• 1259) V1M5 1.5: The laboratory shall record the results obtained, the procedure used for the validation, and a statement as to whether the method is fit for the intended use. The minimum requirements for method validation are given in Sections 1.5.1, 1.5.2 and 1.5.3.

  Yes

  No

  N/A

• 1260) V1M5 1.5: The laboratory shall maintain documentation of the validation procedure for as long as the method is in use and for at least five (5) years past the date of last use.

  Yes

  No

  N/A

• 1261) V1M5 1.5: Laboratories shall participate in a proficiency test program when available. The results of these analyses shall be used to evaluate the ability of the laboratory to produce acceptable data.

  Yes

  No

  N/A

• 1262) V1M5 1.5: The following assessment shall be performed. If no reference method exists, or if the data quality objectives are different from the reference method, then the laboratory shall demonstrate that the method meets the quality objectives for the intended use.

  Yes

  No

  N/A

• 1263) V1M5 1.5.1: Accuracy – Use at least one (1) known pure reference culture at the anticipated environmental conditions, and compare the method results to that of a reference method.

  Yes

  No

  N/A

• 1264) V1M5 1.5.2: Precision – Perform at least ten (10) replicate analyses with both the proposed and reference method, using the target microorganisms of choice. The results shall show that the methods are not statistically different.

  Yes

  No

  N/A

• 1265) V1M5 1.5.3: Selectivity (sensitivity) – Verify all responses in at least ten (10) samples using mixed cultures that include the target organism(s), and at varying concentrations (microbial identification testing or equivalent processes may be used). Calculate the number of false positive and false negative results.

  Yes

  No

  N/A

1266) V1M5 1.6: Demonstration of Capability (DOC)

• 1267) V1M5 1.6.1: General

• 1268) V1M5 1.6.1: Prior to acceptance and institution of any method for data reporting, satisfactory initial DOC is required (see Section 1.6.2).

• 1269) V1M5 1.6.1: Thereafter, ongoing DOC (Section 1.6.3), as per the quality control requirements in Section 1.7.3, is required.

  Yes

  No

  N/A

• 1270) V1M5 1.6.1: In cases where a laboratory analyzes samples using a method that has been in use by the laboratory for at least one year prior to applying for accreditation, and there have been no significant changes in instrument type, personnel or method, the ongoing DOC shall be acceptable as an initial DOC.

  Yes

  No

  N/A

• 1271) V1M5 1.6.1: The laboratory shall have records on file to demonstrate that an initial DOC is not required.

  Yes

  No

  N/A

• 1272) V1M5 1.6.1: For the initial DOC, appropriate records as discussed in Section 1.6.2 shall be completed (1.6.2).

• 1273) V1M5 1.6.1: An initial DOC shall be completed each time there is a change in instrument type, personnel, or method.(1.6.2)

• 1274) V1M5 1.6.1: All demonstrations shall be documented.

  Yes

  No

  N/A

• 1275) V1M5 1.6.1: All data applicable to the demonstration shall be retained and readily available at the laboratory.

  Yes

  No

  N/A

• 1276) V1M5 1.6.2: Initial DOC

• 1277) V1M5 1.6.2: An initial DOC shall be made prior to using any method,

  Yes

  No

  N/A

• 1278) V1M5 1.6.2: An initial DOC shall be at any time there is a change in instrument type, personnel or method

  Yes

  No

  N/A

• 1279) V1M5 1.6.2: An initial DOC shall be made any time that a method has not been performed by the laboratory or analyst in a twelve (12) month period.

  Yes

  No

  N/A

• 1280) V1M5 1.6.2.1: The laboratory shall document each initial DOC in a manner such that the following information is readily available for each affected employee:

  Yes

  No

  N/A

• 1281) V1M5 1.6.2.1 a): analyst(s) involved in preparation and/or analysis;

  Yes

  No

  N/A

• 1282) V1M5 1.6.2.1 b): matrix;

  Yes

  No

  N/A

• 1283) V1M5 1.6.2.1 c): organism(s);

  Yes

  No

  N/A

• 1284) V1M5 1.6.2.1 d): identification of method(s) performed;

  Yes

  No

  N/A

• 1285) V1M5 1.6.2.1 e): identification of laboratory-specific SOP used for analysis, including revision number;

  Yes

  No

  N/A

• 1286) V1M5 1.6.2.1 f): date(s) of analysis;

  Yes

  No

  N/A

• 1287) V1M5 1.6.2.1 g): summary of analyses, including information outlined in Section 1.6.2.2.c.

  Yes

  No

  N/A

• 1288) V1M5 1.6.2.2: If the method or regulation does not specify an initial DOC, the following procedure is acceptable. It is the responsibility of the laboratory to document that other approaches to initial DOC are adequate.

  Yes

  No

  N/A

• 1289) V1M5 1.6.2.2 a): The target organism(s) shall be diluted in a volume of clean quality system matrix (a sample in which no target organisms or interferences are present at concentrations that will impact the results of a specific method).

  Yes

  No

  N/A

• 1290) V1M5 1.6.2.2 a): This matrix shall be sterile phosphate or sterile peptone solution unless specified by the manufacturer.

  Yes

  No

  N/A

• 1291) V1M5 1.6.2.2 a): Prepare at least four (4) aliquots at the concentration specified, or if unspecified, to the countable range for plate methods or working range for most probable number (MPN) type methods.

  Yes

  No

  N/A

• 1292) V1M5 1.6.2.2 b): At least four (4) aliquots shall be prepared and analyzed according to the method either concurrently or over a period of days.

  Yes

  No

  N/A

• 1293) V1M5 1.6.2.2 c): Using all of the results, convert these results to logarithmic values, then calculate the mean recovery and standard deviation of the log converted results in the appropriate reporting units for each organism of interest. When it is not possible to determine mean and standard deviations, such as for presence/absence, the laboratory shall assess performance against established and documented criteria.

  Yes

  No

  N/A

• 1294) V1M5 1.6.2.2 d): For qualitative tests, acceptable performance in a blind study, either internally or externally generated, may be used to meet this Standard, provided that the study consists of a minimum of a blank, a negative culture, and a positive culture for each target organism or metabolite (e.g. b-glucuronidase in E. coli.).

  Yes

  No

  N/A

• 1295) V1M5 1.6.2.2 e): Compare the information from c) above to the corresponding acceptance criteria for precision and accuracy in the method (if applicable or in laboratory-generated acceptance criteria (if there are not established mandatory criteria). If all parameters meet the acceptance criteria, the analysis of actual samples may begin. If any one of the parameters does not meet the acceptance criteria, the performance is unacceptable for that parameter.

  Yes

  No

  N/A

• 1296) V1M5 1.6.2.2 f): When one or more of the tested parameters fail at least one of the acceptance criteria, the analyst shall proceed according to i) or ii) below.

  Yes

  No

  N/A

• 1297) V1M5 1.6.2.2 f) i.: Locate and correct the source of the problem and repeat the initial DOC for all parameters of interest beginning with b) above.

  Yes

  No

  N/A

• 1298) V1M5 1.6.2.2 f) ii.: Repeat the initial DOC for all parameters that failed to meet criteria.

  Yes

  No

  N/A

• 1299) V1M5 1.6.2.2 g): Repeated failure, however, confirms a general problem with the measurement system. If this occurs, locate and correct the source of the problem and repeat the test for all compounds of interest beginning with b).

  Yes

  No

  N/A

• 1300) V1M5 1.6.3: Ongoing DOC

• 1301) V1M5 1.6.3.1: The laboratory shall have a documented procedure describing ongoing DOC.

  Yes

  No

  N/A

• 1302) V1M5 1.6.3.1: The analyst(s) shall demonstrate ongoing capability by meeting the quality control requirements of the method, laboratory SOP, client specifications, and/or this Standard.

  Yes

  No

  N/A

• 1303) V1M5 1.6.3.1: It is the responsibility of the laboratory to document that other approaches to ongoing DOC are adequate.

  Yes

  No

  N/A

• 1304) V1M5 1.6.3.2: This ongoing demonstration may include one of the following or by performing another initial DOC.

• 1305) V1M5 1.6.3.2 a): Analysis of one sample or clean matrix that is fortified with a known quantity of the target organism, with results meeting the laboratory acceptance criteria for accuracy and, where applicable to the testing technique, also meeting the observational details expected for the presumptive, confirmed and completed phases defined in the method.

  Yes

  No

  N/A

• 1306) V1M5 1.6.3.2 b): Analysis of one sample in duplicate for each target organism and test, with results meeting the laboratory acceptance criterion for precision.

  Yes

  No

  N/A

• 1307) V1M5 1.6.3.2 c): Acceptable results for one-single-blind proficiency test sample for target organisms in each field of accreditation.

  Yes

  No

  N/A

• 1308) V1M5 1.6.3.2 d): Performance of an alternate adequate procedure for the field of accreditation, the procedure and acceptance criteria being documented in the laboratory’s quality system.

  Yes

  No

  N/A

• 1309) V1M5 1.6.3.2 e): A documented process of analyst review using QC samples. QC samples can be reviewed to identify patterns for individuals or groups of analysts and determine if corrective action or retraining is necessary; or

  Yes

  No

  N/A

• 1310) V1M5 1.6.3.2 f): f a) through e) are not technically feasible, then analysis of real-world samples with results within a predefined acceptance criteria (as defined by the laboratory or method) shall be performed.

  Yes

  No

  N/A

• 1311) V1M5 1.7: Technical Requirements

1312) V1M5 1.7.1: Calibration

• 1313) V1M5 1.7.1 a): The laboratory shall have documented procedures for calibration, verification, and quality control of support equipment including conductivity meters, oxygen meters, pH meters, hygrometers, and other similar measurement instruments.

  Yes

  No

  N/A

• 1314) V1M5 1.7.1 a): These procedures shall refer to applicable reference methods.

  Yes

  No

  N/A

• 1315) V1M5 1.7.1 b): For instruments that are continuous monitors, such as in-line specific conductance meters:

• 1316) V1M5 1.7.1 b) i.: The laboratory shall document acceptable calibration verification at least once a month.

  Yes

  No

  N/A

• 1317) V1M5 1.7.1 b) ii.: An initial calibration shall be performed if a continuing calibration is unacceptable,

  Yes

  No

  N/A

• 1318) V1M5 1.7.1 b) ii.: An initial calibration shall be performed when the instrument is being returned to service after having been taken off line.

  Yes

  No

  N/A

1319) V1M5 1.7.2: Continuing Calibration

• 1320) V1M5 1.7.2: Reserved for specific procedures.

1321) V1M5 1.7.3: Quality Control

• 1322) V1M5 1.7.3.1: Sterility Checks and Method Blanks

• 1323) V1M5 1.7.3.1 a): Method Blanks

• 1324) V1M5 1.7.3.1 a): The laboratory shall demonstrate that the filtration equipment and filters, sample containers, media and reagents have not been contaminated through improper handling or preparation, inadequate sterilization, or environmental exposure.

  Yes

  No

  N/A

• 1325) V1M5 1.7.3.1 a) i.: For filtration technique, the laboratory shall conduct method blanks per the analytical method.

  Yes

  No

  N/A

• 1326) V1M5 1.7.3.1 a) i.: At a minimum, the filtration series shall include a beginning and ending blank.

  Yes

  No

  N/A

• 1327) V1M5 1.7.3.1 a) i.: The filtration series may include single or multiple filtration units, which have been sterilized prior to beginning the series.

  Yes

  No

  N/A

• 1328) V1M5 1.7.3.1 a) ii.: The filtration series is considered ended when more than thirty (30) minutes elapses between successive filtrations.

  Yes

  No

  N/A

• 1329) V1M5 1.7.3.1 a) ii.: During a filtration series, filter funnels shall be rinsed with three (3) 20-30 ml portions of sterile rinse water after each sample filtration.

  Yes

  No

  N/A

• 1330) V1M5 1.7.3.1 a) ii.: In addition, laboratories shall insert a method blank after every ten (10) samples or sanitize filtration units by UV light after each sample filtration.

  Yes

  No

  N/A

• 1331) V1M5 1.7.3.1 a) iii.: For pour plate technique, method blanks of the medium shall be made by pouring, at a minimum, one uninoculated plate for each lot of pre-prepared, ready-to-use media

  Yes

  No

  N/A

• 1332) V1M5 1.7.3.1 b): Sterility Checks

• 1333) V1M5 1.7.3.1 b) i.: A sterility check shall be analyzed for each lot of pre-prepared, ready-to-use medium (including chromofluorogenic reagent)

  Yes

  No

  N/A

• 1334) V1M5 1.7.3.1 b) i.: A sterility check shall be analyzed for each batch of medium prepared in the laboratory.

  Yes

  No

  N/A

• 1335) V1M5 1.7.3.1 b) i.: A sterility check shall be done prior to first use of the medium.

  Yes

  No

  N/A

• 1336) V1M5 1.7.3.1 b) ii.: For pre-sterilized single use funnels, a sterility check shall be performed on one funnel per lot

  Yes

  No

  N/A

• 1337) V1M5 1.7.3.1 b) ii.: For laboratory-sterilized funnels, a sterility check shall be performed on one funnel per sterilization batch.

  Yes

  No

  N/A

• 1338) V1M5 1.7.3.1 b) iii.: Sterility checks on sample containers shall be performed on at least one (1) container for each lot of purchased, pre-sterilized containers.

  Yes

  No

  N/A

• 1339) V1M5 1.7.3.1 b) iii.: For containers prepared and sterilized in the laboratory, a sterility check shall be performed on one (1) container per sterilized batch with nonselective growth media.

  Yes

  No

  N/A

• 1340) V1M5 1.7.3.1 b) iii.: These sterility checks may be performed by a contracted laboratory if the laboratory does not have the requisite equipment to perform them.

  Yes

  No

  N/A

• 1341) V1M5 1.7.3.1 b) iii.: All correspondence and results from a contracted laboratory shall be retained for a period of five (5) years after the completion of the test(s).

  Yes

  No

  N/A

• 1342) V1M5 1.7.3.1 b) iv.: A sterility check shall be performed on each batch of dilution water prepared in the laboratory with non-selective growth media.

  Yes

  No

  N/A

• 1343) V1M5 1.7.3.1 b) iv.: A sterility check shall be performed on each lot of pre-prepared, ready-to-use dilution water with non-selective growth media.

  Yes

  No

  N/A

• 1344) V1M5 1.7.3.1 b) v.: At least one (1) filter from each new lot of membrane filters shall be checked for sterility with nonselective growth media.

  Yes

  No

  N/A

• 1345) V1M5 1.7.3.2: Test Variability/Reproducibility

• 1347) V1M5 1.7.3.3: Sample Specific Controls (where applicable)

• 1348) V1M5 1.7.3.3 a): Matrix spikes shall be performed per method requirements.

  Yes

  No

  N/A

• 1349) V1M5 1.7.3.3 b): Sample matrix duplicates shall be performed per method requirements.

  Yes

  No

  N/A

• 1350) V1M5 1.7.3.4: Data Reduction

• 1351) V1M5 1.7.3.4: The calculations, data reduction and statistical interpretations specified by each method shall be identified and followed.

  Yes

  No

  N/A

• 1352) V1M5 1.7.3.5: Quality of Standards, Reagents and Media

• 1353) V1M5 1.7.3.5: The laboratory shall ensure that the quality of the reagents and media used is appropriate for the test concerned.

  Yes

  No

  N/A

• 1354) V1M5 1.7.3.5 a): Media – Culture media may be prepared from commercial dehydrated powders or may be purchased ready-to-use.

• 1355) V1M5 1.7.3.5 a) i.: Laboratory-prepared media

• 1356) V1M5 1.7.3.5 a) i. 1: Media prepared by the laboratory from basic ingredients shall be tested for performance (e.g., for selectivity, sensitivity, sterility, growth promotion, and growth inhibition) prior to first use.

  Yes

  No

  N/A

• 1357) V1M5 1.7.3.5 a) i. 2: Media shall be used within the holding time limits specified in the accredited method.

  Yes

  No

  N/A

• 1358) V1M5 1.7.3.5 a) i. 3: Detailed testing criteria information shall be defined in the laboratory’s methods, SOPs, or similar documentation.

  Yes

  No

  N/A

• 1359) V1M5 1.7.3.5 a) ii.: Ready-to-use media

• 1360) V1M5 1.7.3.5 a) ii. 1: Ready-to-use media shall be used within the manufacturer’s expiration date. If the manufacturer’s expiration date is greater than those noted in Section 1.7.3.5 a) i) 2. above, the laboratory shall request, and have available documentation from the manufacturer demonstrating media quality for the extended time period.

  Yes

  No

  N/A

• 1361) V1M5 1.7.3.5 a) ii. 2: Any ready-to-use media used past the expiration date shall be verified for usability by running quality control checks comparing the media with freshly prepared media or by testing recovery with known densities of culture controls.

  Yes

  No

  N/A

• 1362) V1M5 1.7.3.5 b): Reagents and commercial dehydrated powders shall be used within the shelf life of the product, and shall be documented as per TNI Volume 1, Module 2 Quality Systems General Requirements.

  Yes

  No

  N/A

• 1363) V1M5 1.7.3.5 c): Reagent Water

• 1364) V1M5 1.7.3.5 c) i.: The quality of the reagent water used in the laboratory, such as distilled water, de-ionized water or reverse-osmosis produced water shall be monitored for bactericidal and inhibitory substances and shall be used in the preparation of media, solutions and buffers.

  Yes

  No

  N/A

• 1365) V1M5 1.7.3.5 c) ii.: The quality of the water shall be monitored for chlorine residual, specific conductance, total organic carbon, ammonia/organic nitrogen and heterotrophic bacteria plate count monthly (when in use), when maintenance is performed on the water treatment system, or at startup after a period of disuse longer than one month.

  Yes

  No

  N/A

• 1366) V1M5 1.7.3.5 c) iii.: Analysis for metals and the Bacteriological Water Quality Test (to determine presence of toxic agents or growth promoting substances) shall be performed annually. (An exception to performing the Bacteriological Water Quality Test shall be given to laboratories that can supply documentation to show that their water source meets the criteria, as specified by the method, for Type I or Type II reagent water.)

  Yes

  No

  N/A

• 1367) V1M5 1.7.3.5 c) iv.: Results of the above analyses (metals and bacteriological water quality) shall meet the specifications of the required method and records of analyses shall be maintained for five (5) years.

  Yes

  No

  N/A

• 1368) V1M5 1.7.3.5 c) v.: Reagent water purchased from an outside source and used for the preparations of media, solutions and buffers shall meet the criteria specified in items ii) and iii) above. The laboratory shall have documented records of this information. Purchased reagent water that has been in use for longer than the testing intervals specified in items i) through i or in the accredited method shall either be re-tested or discarded.

  Yes

  No

  N/A

• 1369) V1M5 1.7.3.5 d): Documentation for media prepared in the laboratory shall include date of preparation, preparer’s initials, type , manufacturer, lot number, final pH, expiration date, and the amount of reagents used. Documentation for media purchased pre-prepared, ready-to-use (including reagent water purchased from outside sources) shall include manufacturer, lot number, type of media received, date of receipt, expiration date of the media, and pH of the media.

  Yes

  No

  N/A

• 1370) V1M5 1.7.3.6: Selectivity

• 1371) V1M5 1.7.3.6 a): All growth and recovery media shall be checked to assure that the target organism(s) respond in an acceptable and predictable manner.

  Yes

  No

  N/A

• 1372) V1M5 1.7.3.6 b): To ensure that analysis results are accurate, target organism identity shall be verified as specified in the method (e.g., by use of the completed test, or by use of secondary verification tests such as a catalase test or by the use of a completed test such as brilliant green (BG) or E. coli (EC) broth.

  Yes

  No

  N/A

• 1373) V1M5 1.7.3.6 c): In order to ensure identity and traceability, reference cultures used for positive and negative controls shall be obtained from a recognized national collection, organization, or manufacturer recognized by the accreditation body.

  Yes

  No

  N/A

• 1374) V1M5 1.7.3.6 c): Microorganisms may be single use preparations or cultures maintained for their intended use by documented procedures that demonstrate the continued purity and viability of the organism.

  Yes

  No

  N/A

• 1375) V1M5 1.7.3.6 c) i.: Reference cultures may be revived (if freeze-dried) or transferred from slants and sub-cultured once to provide reference stocks. The reference stocks shall be preserved by a technique that maintains the characteristics of the strains. Reference stocks shall be used to prepare working stocks for routine work. If reference stocks have been thawed, they shall not be refrozen and re-used.

  Yes

  No

  N/A

• 1376) V1M5 1.7.3.6 c) ii.: Working stocks shall not be sequentially cultured more than five (5) times and shall not be sub-cultured to replace reference stocks.

  Yes

  No

  N/A

• 1377) V1M5 1.7.3.6 d): Culture Controls

• 1378) V1M5 1.7.3.6 d) i.: Negative Culture Controls

• 1379) V1M5 1.7.3.6 d) i. 1: Negative culture controls demonstrate that the medium does not support the growth of non-target organisms or does not exhibit the typical positive reaction of the target organism(s).

• 1380) V1M5 1.7.3.6 d) i. 2: Each pre-prepared, ready-to-use lot of selective medium (including chromofluorogenic reagent) and each batch of selective medium prepared in the laboratory shall be analyzed with one or more known negative culture controls (i.e. non-target organisms), as appropriate to the method. This shall be done prior to first use of the medium.

  Yes

  No

  N/A

• 1381) V1M5 1.7.3.6 d) ii.: Positive Culture Controls

• 1382) V1M5 1.7.3.6 d) ii. 1: Positive culture controls demonstrate that the medium can support the growth of the target organism(s), and that the medium produces the specified or expected reaction to the target organism(s).

• 1383) V1M5 1.7.3.6 d) ii. 2: Each pre-prepared, ready-to-use lot of medium (including chromofluorogenic reagent) and each batch of medium prepared in the laboratory shall be tested with at least one pure culture of a known positive reaction. This shall be done prior to first use of the medium.

  Yes

  No

  N/A

• 1384) V1M5 1.7.3.7: Constant and Consistent Test Conditions

• 1385) V1M5 1.7.3.7 a): Laboratory Facilities

• 1386) V1M5 1.7.3.7 a): Floors and work surfaces shall be non-absorbent and easy to clean and disinfect. Work surfaces shall be adequately sealed.

  Yes

  No

  N/A

• 1387) V1M5 1.7.3.7 a): Laboratories shall provide sufficient storage space, and shall be clean and free from dust accumulation.

  Yes

  No

  N/A

• 1388) V1M5 1.7.3.7 a): Plants, food, and drink shall be prohibited from the laboratory work area.

  Yes

  No

  N/A

• 1389) V1M5 1.7.3.7 b): Laboratory Equipment

• 1390) V1M5 1.7.3.7 b) i.: Temperature Measuring Devices

• 1391) V1M5 1.7.3.7 b) i.: Temperature measuring devices such as liquid-in-glass thermometers, thermocouples, and platinum resistance thermometers used in incubators, autoclaves and other equipment shall be the appropriate quality to meet specification(s) in the method.

  Yes

  No

  N/A

• 1392) V1M5 1.7.3.7 b) i.: The graduation of the temperature measuring devices shall be appropriate for the required accuracy of measurement and they shall be verified to national or international standards for temperature.

  Yes

  No

  N/A

• 1393) V1M5 1.7.3.7 b) i.: Verification shall be done at least annually (see TNI Volume 1, Module 2, Section 5.5.13.1).

  Yes

  No

  N/A

• 1394) V1M5 1.7.3.7 b) ii.: Autoclaves

• 1395) V1M5 1.7.3.7 b) ii.: The performance of each autoclave shall be initially evaluated by establishing its functional properties and performance, for example heat distribution characteristics with respect to typical uses.

  Yes

  No

  N/A

• 1396) V1M5 1.7.3.7 b) ii.: Autoclaves shall meet specified temperature tolerances. Pressure cookers shall not be used for sterilization of growth media.

  Yes

  No

  N/A

• 1397) V1M5 1.7.3.7 b) ii.: Pressure cookers shall not be used for sterilization of growth media.

  Yes

  No

  N/A

• 1398) V1M5 1.7.3.7 b) ii.: Demonstration of sterilization temperature shall be provided by use of a continuous temperature recording device or by use of a maximum registering thermometer with every cycle.

  Yes

  No

  N/A

• 1399) V1M5 1.7.3.7 b) ii.: At least once during each month that the autoclave is used, appropriate biological indicators shall be used to determine effective sterilization. The selected biological indicator shall be effective at the sterilization temperature and time needed to sterilize lactose-based media.

  Yes

  No

  N/A

• 1400) V1M5 1.7.3.7 b) ii.: Temperature sensitive tape shall be used with the contents of each autoclave run to indicate that the autoclave contents have been processed.

  Yes

  No

  N/A

• 1401) V1M5 1.7.3.7 b) ii.: Records of autoclave operations shall be maintained for every cycle. Records shall include: date, contents, maximum temperature reached, pressure, time in sterilization mode, total run time (may be recorded as time in and time out) and analyst’s initials.

  Yes

  No

  N/A

• 1402) V1M5 1.7.3.7 b) ii.: Autoclave maintenance, either internally or by service contract, shall be performed annually, and shall include a pressure check and verification of temperature device. Records of the maintenance shall be maintained in equipment logs.

  Yes

  No

  N/A

• 1403) V1M5 1.7.3.7 b) ii.: NOTE: When it has been determined that the autoclave has no leaks, pressure checks can be documented using the formula PV = nRT.

  Yes

  No

  N/A

• 1404) V1M5 1.7.3.7 b) ii.: The autoclave mechanical timing device shall be checked quarterly against a stopwatch and the actual time elapsed documented.

  Yes

  No

  N/A

• 1405) V1M5 1.7.3.7 b) iii.: Volumetric Equipment

• 1406) V1M5 1.7.3.7 b) iii.: Volumetric equipment shall be verified as follows:

• 1407) V1M5 1.7.3.7 b) iii. 1: equipment with movable parts such as automatic dispensers, dispensers/diluters, and mechanical hand pipettes shall be verified for accuracy quarterly.

  Yes

  No

  N/A

• 1408) V1M5 1.7.3.7 b) iii. 2: equipment such as filter funnels, bottles, non-Class A glassware, and other containers with volumetric markings (including sample analysis vessels) shall be verified once per lot prior to first use. This verification may be volumetric or gravimetric.

  Yes

  No

  N/A

• 1409) V1M5 1.7.3.7 b) iii. 3: the volume of the disposable volumetric equipment such as sample bottles, and disposable pipettes shall be checked once per lot.

  Yes

  No

  N/A

• 1410) V1M5 1.7.3.7 b) iv.: UV Instruments

• 1411) V1M5 1.7.3.7 b) iv.: UV instruments, used for sanitization, shall be tested quarterly for effectiveness with an appropriate UV light meter, by plate count agar spread plates or other methods providing equivalent results such as uvcide strips.

  Yes

  No

  N/A

• 1412) V1M5 1.7.3.7 b) iv.: Replace bulbs if output is less than 70% of original for light tests or if count reduction is less than 99% for a plate containing 200 to 300 organisms.

  Yes

  No

  N/A

• 1413) V1M5 1.7.3.7 b) v.: Incubators, Water Baths, Ovens

• 1414) V1M5 1.7.3.7 b) v. 1: The uniformity of temperature distribution in incubators and water baths shall be established.

  Yes

  No

  N/A

• 1415) V1M5 1.7.3.7 b) v. 2: Records shall be maintained for each cycle that include date, cycle time, temperature, contents and analyst’s initials.

  Yes

  No

  N/A

• 1416) V1M5 1.7.3.7 b) vi.: Labware (Glassware and Plasticware)

• 1417) V1M5 1.7.3.7 b) vi. 1: The laboratory shall have a documented procedure for washing labware, if applicable. Detergents designed for laboratory use shall be used.

  Yes

  No

  N/A

• 1418) V1M5 1.7.3.7 b) vi. 2: Glassware shall be made of borosilicate or other non-corrosive material, free of chips and cracks, and shall have readable measurement marks.

  Yes

  No

  N/A

• 1419) V1M5 1.7.3.7 b) vi. 3: Labware that is washed and reused shall be tested for possible presence of residues that may inhibit or promote growth of microorganisms by performing the Inhibitory Residue Test annually, and each time the lab changes the lot of detergent or washing procedures.

  Yes

  No

  N/A

• 1420) V1M5 1.7.3.7 b) vi. 4: Washed labware shall be tested at least once daily, each day of washing, for possible acid or alkaline residue by testing at least one piece of labware with a suitable pH indicator such as bromothymol blue. Records of tests shall be maintained.

  Yes

  No

  N/A

1421) V1M5 1.7.4: Data Acceptance/Rejection Criteria

• 1422) V1M5 1.7.4: Method criteria and evaluation methods shall be used.

  Yes

  No

  N/A

1423) V1M5 1.7.5: Sample Handling

• 1424) V1M5 1.7.5 a): Samples that require thermal preservation shall be considered acceptable if the arrival temperature of a representative sample container meets the method or mandated temperature requirement.

  Yes

  No

  N/A

• 1425) V1M5 1.7.5 a) i.: Samples that are delivered to the laboratory on the same day they are collected may not meet the requirements of Section 1.7.5.a). In these cases, the samples shall be considered acceptable if the samples were received on ice.

  Yes

  No

  N/A

• 1426) V1M5 1.7.5 a) ii.: If sample analysis is begun within fifteen (15) minutes of collection, thermal preservation is not required.

  Yes

  No

  N/A

• 1427) V1M5 1.7.5 a) iii.: Thermal preservation is not required in the field if the laboratory receives the sample and either begins the analysis or refrigerates the sample within fifteen (15) minutes of collection.

  Yes

  No

  N/A

• 1428) V1M5 1.7.5 b): Microbiological samples from known chlorinated sources (such as wastewater effluent), unknown sources where chlorine usage is suspected (such a new client or a new source) and all potable water sources (including source water) shall be checked for absence of chlorine residual.

  Yes

  No

  N/A

• 1429) V1M5 1.7.5 b): Laboratories that receive samples from potable water sources (including source water) that have a demonstrated history of acceptable preservation may check a sample from each source at a frequency of once per month if:

• 1430) V1M5 1.7.5 b) i.: the laboratory can show that the received sample containers are from their laboratory;

  Yes

  No

  N/A

• 1431) V1M5 1.7.5 b) ii.: sufficient sodium thiosulfate was in each container before sample collection to neutralize at minimum 5 mg/l of chlorine for drinking water and 15 mg/l of chlorine for wastewater samples;

  Yes

  No

  N/A

• 1432) V1M5 1.7.5 b) iii.: one container from each batch of laboratory prepared containers or lot of purchased ready-to-use containers is checked to ensure efficacy of the sodium thiosulfate to 5 mg/l chlorine or 15 mg/l chlorine as appropriate and the check is documented;

  Yes

  No

  N/A

• 1433) 1.7.5 b) iv.: chlorine residual is checked in the field and actual concentration is documented with sample submission.

  Yes

  No

  N/A