Form TBD Checklist A - Method 1623_1623.1 Audit Package and Data

Checklist A – Method 1623/1623.1 Audit Package and Data Review
Laboratory Name

Name and Affiliation of Evaluator

Date of Evaluation

Good Laboratory Practice (GLP) is generally defined as a system of management controls for the laboratories to ensure the consistency and reliability of results. Adapted
from other federal programs for the purposes of the Cryptosporidium Laboratory QA Evaluation Program, GLP includes personnel, equipment, and standard operating
procedures appropriate for the program.

Reference*

Item to be Evaluated
1623

1623.1

Cert

Yes

9.1

9.1

1.

Requirement
GLP

22.2

22.2

1.1 -1.3

Requirement
GLP

9.1

9.1

1.7

Critical
GLP

-

-

1.7

Critical
GLP

10.6

9.10

7.1.9

Requirement

10.6.4

9.10.4

7.1.9.5

Recommendation

1 Quality Assurance
1.1

Is documentation (e.g., resume, sample list)
available for all Method 1623/1623.1 staff?
1.1.1

1.2

Are employee training records available and up to
date?
1.2.1

1.3

1.5

Have all analysts documented that they
have read and understood the QA Plan
and SOPs?

Is the laboratory performing analyst verification
monthly and does the lab have corrective action
procedures in place if criteria are not met?
1.3.1

1.4

Have technicians/analysts analyzed the
required number of samples using
Method1623 or1623.1?

If the laboratory has only one analyst, is
the analyst demonstrating analyst
verification through comparison with
photo libraries or repetitive counts?

Does the quality assurance plan address
requirements for Cryptosporidium analysis under
LT2ESWTR?
Have acceptable initial precision and recovery
analyses been performed for each version of the
method the laboratory is using?

Critical

9.1.2.1.1

9.2

Satisfactory

Classification

7.1.4

G-7

Requirement

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
1.6

Of the field/PT samples reviewed, is each field/PT
sample associated with an acceptable method
blank?

1.7

1623.1

Cert

Yes

9.6.1

9.7

7.1.2
7.1.5.1

Requirement

Were all method blanks (MB) evaluated
without contamination?

9.6.2.1

9.7.2

7.1.5

Requirement

1.6.2

Were the same lots of reagents (elution,
IMS, and staining) used for the method
blank and the associated field/PT
samples?

-

-

7.1.5.3

Critical

9.6

9.7

7.1.5.2

Requirement

9.7

9.8

7.1.2
7.1.6.1

Requirement

9.7.3
Table 3
Table 4

9.8.3
Table 3
Table 4

7.1.6.2

Is method blank analyzed prior to the
analysis of field/PT samples?

Is each field/PT sample associated with an
acceptable ongoing precision and recovery (OPR)
sample?
What percentage of OPR samples
evaluated met the recovery criteria?

1.7.2

Were the same lots of reagents (elution,
IMS, and staining) used for the OPR and
the associated field/PT samples?

-

-

7.1.6.1

Critical

1.7.3

Is OPR analyzed prior to the analysis of
field/PT samples?

9.7.1

9.8.1

7.1.6.2

Requirement

1.7.4

Does the laboratory maintain control
charts of OPR results?

9.7.6
9.1

9.8.3
9.12.1
Table 2

7.1.7

1623
Recommendation

9.4.3
Table 3
Table 4

9.5.3
Table 3
Table 4

7.1.6.2

QC Criteria

9.1.8

9.6.1

7.1.2
7.1.10.2

Requirement

What is the mean and relative standard
deviation (RSD), or standard deviation, of
the recoveries of the OPR samples
included in the control chart?

Were matrix spike (MS) samples analyzed at the
minimum frequency of 1 MS per 20 (up to and
including) field samples from each source?

No

NA

Comments/
Response Requested

UNK

# MB reviewed:

# OPR reviewed:

1.7.1

1.7.5

1.8

1623

1.6.1

1.6.3

Satisfactory

Classification

1623.1 Requirement
Mean:

1.8.1

Were MS sample volumes within 10% of
their associated field samples’ volumes?

9.5.1

9.6.2

7.1.10.3

Requirement

1.8.2

Were MS samples analyzed at the same
time and using the same method
variation as their associated field
samples?

Table 2

Table 2

7.1.10.1

Requirement

G-8

RSD:

# MS reviewed:

Reference*

Item to be Evaluated
1623

1623.1

Cert

What is the mean and relative standard
deviation of the MS samples reviewed?

Table 3
Table 4
Table 5

Table 3
Table 4
Table 6

-

QC Criteria

Does the laboratory maintain control
charts of MS results?

9.5.1.4
9.1

9.6.2.3
9.12.2
Table 2

7.1.10.4

1623
Recommendation

Were OPR samples spiked with 100 - 500
organisms?

9.7

9.8

7.1.6.1

Requirement

1.10 Does the laboratory perform IMS controls and
maintain IMS control charts? If not, how do they
troubleshoot low recoveries?

9.7.5.3

9.8.7.3
9.13

-

Recommendation

1.11 Does the laboratory have an adequate record
system for tracking samples, including unique ID,
from collection through log-in, analysis, and data
reporting?

-

-

8.0

Critical
GLP

1.8.3

1.8.4

1.9

2.1

Yes

1623.1 Requirement

1.12 Is the laboratory using the Method December
2005 version of Method 1623 or Method 1623.1
for LT2 samples?

2

Requirement

Data Recording Procedures
Is shipping information complete, i.e., time/date of
sample collection, sampler's name, time/date of
sample receipt, receiver's initials, sample
condition?
2.1.1

Were all samples evaluated received at
≤20°C and not frozen?

Satisfactory

Classification

8.1.3

8.1.3

8.5

Requirement

8.1.3

8.1.3

6.3.3

Requirement

2.2

Do sample numbers on the chain of custody
match the sample numbers on the report forms?

-

-

-

Requirement

2.3

Are current Method 1623/1623.1 bench sheets
used to record sample processing data?

-

-

8.2

Recommendation

2.4

Are all primary measurements during each step
recorded, including all raw data used in
calculations?

9.1.2.2.5

9.3.5

8.0

Requirement

2.5

Technician/analyst, date, and time of elution is
recorded?

12.2.6.2.1

12.2.7.1
12.3.2.1

8.7

Requirement

2.6

Technician/analyst, date, and time of slide
preparation is recorded?

13.3.3.11

13.3.3.11

8.7

Requirement

G-9

No

NA

Comments/
Response Requested

UNK
Mean:
RSD:

Reference*

Item to be Evaluated
1623

1623.1

Cert

Yes

14.10

14.10

8.7

Requirement

2.7

Technician/analyst, date, and time of staining is
recorded?

2.8

Are batch and lot numbers of reagents used in the
analysis of the sample recorded?

-

-

8.7

Critical

2.8.1

Lot number for the IMS kit is recorded?

-

-

8.7

Critical

2.8.2

Lot number of the staining kit is
recorded?

-

-

8.7

Critical

2.8.3

Lot number of the spiking suspensions is
recorded?

-

-

3.21.4
8.7

Critical

-

-

3.21.4
8.6

Requirement

15.2

15.2

8.2

Requirement

2.11 Name of examining analyst is recorded?

15.2.6

15.2.6

8.7

Requirement

2.12 Date and time of sample examination is recorded?

15.2.4

15.2.4

8.7

Requirement

-

-

-

Requirement

15.2.2.3
15.2.3.3

15.2.2.4
15.2.3.4

5.4.9.4
5.4.10.4

Requirement

2.15 Is each reported positive organism detected in a
field sample characterized and recorded?

15.2

15.2.2.1
15.2.3.1

5.4.9.1
5.4.10.1

Requirement

2.16 Do values recorded on the data sheets match the
values reported to the client?

-

-

8.1

Requirement

2.17 Are mistakes on all forms crossed out with a
single line, initialed, and dated?

-

-

8.2

Critical

2.18 Are data always legible and recorded in pen?

-

-

8.2

Critical

2.19 Was the final report reviewed by QA manager, lab
director or an individual other than the analyst?

-

-

8.1

Critical

2.20 Do records demonstrate each analyst's
characterization of 3 oocysts and 3 cysts from
positive control for each microscopy session?

15.2.1.1

15.2.1.1

5.4.6

Requirement

2.21 Data shows that no more than 0.5 mL of pellet
was used per IMS?

13.2.4

13.2.3

5.2.3
8.6

Requirement

2.9

Spike value recorded for all spiked samples?

2.10 Are Method 1623/1623.1 Cryptosporidium Slide
Examination forms used to record sample
examination results?

2.13 Are calculations of final concentrations and
recoveries complete and correct?
2.14 Is the size of the cysts and oocysts reported to the
nearest 0.5 µm?

Satisfactory

Classification

G-10

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
3

1623

1623.1

Cert

Yes

No

NA

UNK

Comments/
Response Requested

Holding Times –Method 1623.1

3.1

Is sample elution initiated within 96 hours of
sample collection or field filtration?

8.2.1
Table 1

8.2.1
Table 5

6.4
8.7

Requirement

3.2

Are sample elution, concentration, and purification
steps completed in one work day?

8.2.2
Table 1

8.2.2
Table 5

6.4
8.7

Requirement

3.3

Are slides stained within 72 hours of application of
the purified sample to the slide?

8.2.3
Table 1

8.2.3
Table 5

6.4
8.7

Requirement

3.4

Are stained slides read and confirmed within 7
days of staining? [Section 8.2.4 and Table 5]

8.2.4
Table 1

8.2.4
Table 5

6.4
8.7

Requirement

4

Satisfactory

Classification

Spike enumeration procedures

4.1

Source of flow cytometry-enumerated spiking
suspensions.

-

11.2

-

4.2

If 50-L samples are analyzed, what positive
control procedure does the laboratory follow for
OPR and MS samples: (A) spike entire 50 L, (B)
spike and filter 10 L before filtering 40 L, or (C)
filter 40 L before spiking and filtering 10 L.

-

-

7.1.10.3

The following items below are optional if the laboratory is NELAC certified. If the laboratory opts to provide NELAC certification, complete the box below by entering the NELAC certification number and
date. Provide copy of certification.
NELAC Certification Number:

5

Certification Date:

Laboratory Equipment and Supplies

5.1 Reagent-grade water testing
5.1.1

Is reagent water tested monthly for
conductivity and total chlorine residual?

-

-

4.3.1

Critical
GLP

Were the results for the above
parameters acceptable? Total
chlorine residual not greater
than 0.1 mg/L, conductivity not
greater than 2 µmhos/cm?

-

-

4.3.1

Critical
GLP

Has the reagent water been tested
annually for metals – Pb, Cd, Cr, Cu, Ni,
Zn?

-

-

4.3.1

Critical
GLP

5.1.1.1

5.1.2

G-11

Reference*

Item to be Evaluated
1623

1623.1

Cert

-

-

4.3.1

Critical
GLP

-

-

4.3.1

Critical
GLP

-

-

4.3.1

Critical
GLP

Is still or DI unit maintained according to
manufacturer's instructions?

-

-

4.3.3

Critical
GLP

5.2.1

Accuracy ± 0.1 units, scale graduations,
0.1 units?

-

-

3.1.1

Critical
GLP

5.2.2

Is a record maintained for pH
measurements and calibrations?

-

-

3.1.4

Critical
GLP

5.2.3

Is pH meter standardized each use
period with pH 7, 4 or 10 standard buffers
(selection dependent upon desired pH)?

-

-

3.1.4

Critical
GLP

Are all pH buffers dated when received
and opened, and discarded before
expiration date?

-

-

3.1.5

Critical
GLP

Are balance calibrations verified monthly
using ASTM Class 1, Class 2 or Class 3
weights or weights traceable to Class 1,
Class 2, or Class 3 weights, or
equivalent? Non-reference weights
should be calibrated every six months
with reference weights.

-

-

3.2.2

Critical
GLP

5.3.2

Is correction data and Certificate of
Traceability available for weights?

-

-

3.2.3

Critical
GLP

5.3.3

Is preventative maintenance conducted
yearly at a minimum?

-

-

3.2.4

Recommendation
GLP

5.1.2.1

5.1.3

Is reagent water tested monthly for
heterotrophic plate count?
5.1.3.1

5.1.4

Were the results for the metals
testing acceptable; each metal
not greater than 0.05 mg/L and
collectively not greater than 0.1
mg/L?

Are the results for the
heterotrophic plate count
acceptable, < 500 CFU/mL?

Yes

5.2 pH meter

5.2.4

5.3 Balances (top loader or pan balance)
5.3.1

Satisfactory

Classification

5.4 Temperature recording device

G-12

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
1623

1623.1

Cert

Are calibration of thermometers checked
annually (dial thermometers quarterly) at
the temperature used against a reference
NIST thermometer or equivalent?

8.1.4

8.1.4

3.3.2

Requirement
GLP

Is the sample storage refrigerator able to
maintain temperature of 1 to 10°C?

-

-

3.7.1

Critical
GLP

9.2.1

Appendix
A

3.8.2

Requirement
GLP

9.1

9.1

3.15.5

Critical
GLP

-

-

3.15.5

Critical
GLP

Are date, contents, sterilization time and
temperature, and technician initials
recorded for each cycle?

-

-

3.5.3

Critical
GLP

Is a maximum registering thermometer or
continuous monitoring device used
during each autoclave cycle?

-

-

3.5.5

Critical
GLP

5.7.3

Is automatic timing mechanism checked
with stopwatch quarterly?

-

-

3.5.6

Critical
GLP

5.7.4

Are spore strips or ampules used
monthly to confirm sterilization?

-

-

3.5.5

Critical
GLP

9.1

9.1

4.5.1

Requirement

4.5.2

Recommendation

4.5.2

Recommendation
GLP

5.4.1

5.4.2

Yes

5.5 Micropipetters
5.5.1

Have micropipetters been calibrated
within the past year?

5.6 Centrifuge
5.6.1

Is a maintenance contract in place or
internal maintenance protocol available?

5.6.2

Is the centrifuge calibrated yearly?

5.7 Autoclave
5.7.1

5.7.2

6

Quality Assurance Manual

6.1

Does the laboratory have a formal QA laboratory
plan prepared and ready for examination?

6.2

Is a laboratory organization chart or other
information available listing staff organization and
responsibilities? Does it identify the QA manager
and lab director?
6.2.1

Is the QA manager separate from the lab
director?

-

-

Satisfactory

Classification

G-13

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
6.3

Does the laboratory have a schedule and/or
procedure for all preventative maintenance of
equipment?

Satisfactory

Classification

1623

1623.1

Cert

-

-

4.5.3

Comments:

G-14

Yes
GLP

No

NA

UNK

Comments/
Response Requested