TBD Checklist B - Laboratory SOP Review

Checklist B - Laboratory SOP 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
For each item, does the SOP specify:
1

1623

1623.1

Cert

Yes

Sample Spiking

1.1

The suspension vial is vortexed for 30 seconds or per
11.4.3.1.2
manufacturer’s instructions?

1.2

The carboy used for the method blank is randomly
selected from carboy stock to check efficacy of
cleaning system or disposable carboys are used for
all samples?

1.3

The details of the suspension vial rinse, including
volumes?

1.4

Acceptable sample spiking procedures, including
issues not noted in items 1.1 through 1.3?

2

Satisfactory
Classification

11.2.3.2

-

Method Procedure

-

-

7.1.5.3

Critical

11.4.3.1

11.2.3

-

Method Procedure
Critical
GLP

Filtration/Elution

2.1 Envirochek® HV filtration
2.1.1

The flow rate is maintained at
approximately 2 L/min?

12.2.1.2

12.2.1.2

-

Method Procedure

2.1.2

The volume filtered is measured using a
flow totalizer or calibrated carboy?

12.2.4.2

12.2.4.2

-

Requirement

2.1.3

The sample is stirred during filtration?

12.2.4.1

12.2.4.1

-

Method Procedure

2.1.4

The details of the carboy rinse after
filtration including volume?

12.2.4.5

12.2.4.6

-

Method Procedure

G-15

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
For each item, does the SOP specify:
2.1.5

Appropriate maintenance and cleaning
procedures?

2.1.6

Acceptable Envirochek® filtration
procedures, including issues not noted in
items 2.1.1 through 2.1.5?

Satisfactory
Classification

1623
-

1623.1
-

Cert

Yes

-

Critical
Critical
GLP

2.2 Envirochek® HV capsule filter elution
2.2.1

Measurement of the volume of the elution
buffer used or that the volume covers the
membrane?

12.2.6.2.2

12.2.8.2

-

Method Procedure

2.2.2

The speed that samples are shaken?

12.2.6.2.3

12.2.8.3

-

Method Procedure

2.2.3

The dispersant is added to the sample as
per Method 1623.1?

12.2.7

-

1623
Recommendation

2.2.4

1623.1 Requirement

The samples are shaken three times for 5
minutes each time, and each in a different
orientation?

12.2.6.2

12.2.8

-

Method Procedure

2.2.5

Procedures for filter capsule rinse and
addition of rinsate to the centrifuge bottle?

12.2.6.2.8

12.2.8.8

-

Method Procedure

2.2.6

Acceptable Envirochek® capsule filter
elution procedures, including issues not
noted in items 2.2.1 through 2.2.5?

Critical
GLP

2.3 Filta-Max® filtration
2.3.1

The flow rate is maintained at ≤4 L per
minute for Filta-Max®?

12.3.1.1.3

12.3.1.1.3

-

Method Procedure

2.3.2

The volume filtered is measured using a
flow totalizer or calibrated carboy?

12.3.1.5.2

12.3.1.5.2

-

Requirement

2.3.3

Appropriate maintenance and cleaning
procedures? [Section 12.3.4]

12.3.4

12.3.4

-

Requirement

2.3.4

Acceptable Filta-Max® filtration
procedures, including issues not noted in
items 2.3.1 through 2.3.3?

Critical
GLP

2.4 Filta-Max® filter wash station elution
2.4.1

The use of PBST to elute the filter?

7.4.2.4

7.6.2.4

-

Method Procedure

G-16

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
For each item, does the SOP specify:
2.4.2

The amount of PBST used for each wash?
(approx. 600 mL)

2.4.3

1623

1623.1

Cert

Yes

12.3.2.2

12.3.2.2

-

Method Procedure

The plunger is moved up and down 20
times during the first wash?

12.3.2.2.1
h

12.3.2.2.1
h

-

Method Procedure

2.4.4

The plunger is moved up and down gently
to avoid generating excess foam?

12.3.2.2.1
h

12.3.2.2.1
h

-

Method Procedure

2.4.5

That during the second wash the plunger
is moved up and down 10 times?

12.3.2.2.2
b

12.3.2.2.2
b

-

Method Procedure

2.4.6

The instructions for cleaning the wash
station between samples?

12.3.4.2

12.3.4.2

-

Requirement

2.4.7

The housing is rinsed after filter is
removed and the rinse is included in the
sample volume?

12.3.2.2.1
d

12.3.2.2.1
d

-

Method Procedure

2.4.8

3

Satisfactory
Classification

Acceptable Filta-Max® filter wash station
elution procedures, including issues not
noted in items 2.4.1 through 2.4.7?

Critical
GLP

Concentration

3.1 Filta-Max® filter sample concentration (as an alternative or in addition to Section 3.2)
3.1.1

The force of the vacuum is maintained
below 30 cm Hg?

3.1.2
3.1.3

3.1.4

NOTE
pg 43

NOTE
pg 34

-

Method Procedure

That concentration is performed after each
of the washes?

12.3.2.2.1
j

12.3.2.2.1
j

-

Method Procedure

The sample is concentrated so that some
liquid remains above the filter (enough to
cover the stir bar about half-way)?

12.3.3.2.1
c

12.3.3.2.1
b

-

Method Procedure

12.3.3.2

12.3.3.2

-

Requirement

12.3.3.2.3

12.3.3.2.3

-

Method Procedure

The stir bar and concentration tube are
rinsed after each concentration and the
liquid added to the concentrate?

3.1.5

The filter membrane is washed twice with
5 mL of PBST each time?

3.1.6

Acceptable Filta-Max® filter sample
concentration procedures, including issues
not noted in items 3.1.1 through 3.1.5?

Critical
GLP

3.2 Envirochek® HV and Filta-Max® filter sample centrifugation

G-17

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
For each item, does the SOP specify:

1623.1

Cert

Yes

13.2.1
including
NOTE

-

Method Procedure

Instructions to ensure the centrifuge tubes
are properly balanced prior to
centrifugation?

-

-

3.15.4

Critical

The sample is centrifuged for 15 minutes
with start time beginning when centrifuge
reaches the required speed?

13.2.1

13.2.1

-

Method Procedure

3.2.4

The centrifuge is slowly decelerated at the
end without using the brake?

13.2.1

13.2.1

-

Method Procedure

3.2.5

Acceptable Envirochek® HV and FiltaMax® filter sample centrifugation
procedures, including issues not noted in
items 3.2.1 through 3.2.4?

3.2.2

3.2.3

4.1

1623
13.2.1
including
NOTE

3.2.1

4

Satisfactory
Classification

The sample is centrifuged at 1500 x G
(maximum 2000 x G) using a swinging
bucket rotor?

Critical
GLP

Purification and Slide Preparation
The centrifuged sample supernatant is aspirated no
lower than 5 mL of supernatant above every 0.5 mL
of the pellet or portion of 0.5 mL pellet?

13.2.2

13.2.2
13.2.3

5.2.2
5.2.3

Requirement

4.1.1

The type and internal diameter of pipette
used for aspiration of supernatant?

-

NOTE
pg 37

-

Recommendation

4.1.2

The rate of aspiration (i.e., mL/ min or
pressure of the vacuum)?

-

13.2.2

-

Recommendation

4.2

The tube is vortexed vigorously until pellet is
completely resuspended?

13.2.3

13.2.2.1

-

Method Procedure

4.3

Appropriate procedures for dividing pellets greater
than 0.5 mL into subsamples and the analysis of
the subsamples?

13.2.4

13.2.3

-

Critical

4.4

No more than 0.5 mL of pellet is used per IMS?

13.2.4

13.2.3

5.2.3

Method Procedure

4.5

The resuspended pellet volume is quantitatively
transferred to the flat-sided tube (2 rinses) including
the determination of the rinse volumes?

13.3.2.1

13.3.2.1

-

Method Procedure

4.6

SL-Buffer A is used at room temperature or that it is
checked for precipitate before use?

NOTE
pg 47

NOTE
pg 39

3.17.2

Method Procedure

4.7

The volume of 10x SL-Buffer A is 1 mL?

13.3.1.2

13.3.1.2

5.2.5

Method Procedure

G-18

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
For each item, does the SOP specify:

Satisfactory
Classification

1623

1623.1

Cert

Yes

4.8

The volume of 10x SL-Buffer B is 1 mL?

13.3.1.3

13.3.1.3

5.2.5

Method Procedure

4.9

Instructions for thorough resuspension of IMS
beads prior to addition to the flat-sided tube?

13.3.2.2
13.3.2.4

13.3.2.2
13.3.2.4

-

Method Procedure

4.10 100 µL of Cryptosporidium and Giardia beads are
used?

13.3.2.3
13.3.2.5

13.3.2.3
13.3.2.5

5.2.5

Method Procedure

4.11 The flat-sided tube is rotated at 18 rpm for 1 hour at
room temperature?

13.3.2.6

13.3.2.6

-

Method Procedure

4.12 Which magnetic concentrators, MPC®-1 or MPC®-6,
are used?

Method Procedure

4.13 The placement of the flat-sided tube in the magnet
and the rock technique and time?

13.3.2.9

13.3.2.8
13.3.2.9

-

Method Procedure

4.14 The sample is quantitatively transferred from the
flat-sided tube to the microcentrifuge tube (2 rinses)
including rinse volumes?

13.3.2.13

13.3.2.14

-

Method Procedure

4.15 The flat-sided tube is allowed to sit one minute after
each transfer to accumulate residual sample, then
the residual is transferred to microcentrifuge tube?

13.3.2.13

13.3.2.14

-

Method Procedure

4.16 The magnet is in the vertical position in the MPC®S?

-

13.3.2.13

-

Method Procedure

13.3.4

13.3.2.17

-

1623
Recommendation

4.17 The beads are rinsed with PBS while inside the
microcentrifuge tube?
4.18 Standard NaOH (5 µL, 1N) and standard HCl (50
µL, 0.1N) are used?

1623.1 Requirement
NOTES
pg 49-50

NOTES
pg 42

3.17.5

Requirement

4.19 The sample is vortexed vigorously for 50 seconds
immediately after the addition of acid and 30
seconds after the sample has set for 10 minutes at
room temperature?

13.3.3

13.3.3

-

Method Procedure

4.20 The magnet is in the slanted position in the MPC®-S
for dissociation steps?

-

13.3.3.6

-

Method Procedure

13.3.3.10

13.3.3.10

5.2.4

Requirement

4.21 A second dissociation is performed?

G-19

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
For each item, does the SOP specify:

1623

4.22 When the second dissociation is performed, the
laboratory:
A) uses a second slide, or

Satisfactory
Classification

13.3.3.10

1623.1

13.3.3.10
13.4.5

Cert

Yes

Circle one:

-

A

B

B) adds the additional volume to the original slide?
4.23 The volume and the timing of the NaOH addition to
the wells?

13.3.3.8

13.3.3.8

-

Method Procedure

13.3.3.12

13.3.3.12

-

4.25 If the laboratory has more than one option specified
for slide drying, are criteria included for when each
option will be used?

-

-

5.3.1

Recommendation

4.26 That positive and negative staining controls are
prepared at the same time the slides are prepared?

14.1

14.1.3

-

Requirement

4.24 When the slides are dried (e.g., room temperature
or slide warmer), the laboratory:
A) uses room temperature, or
o

Circle one:
A

o

B) uses 35 to 42 C, or

B

C

C) follows manufacturer’s instructions?

4.27 Acceptable sample purification and slide
preparation procedures, including issues not noted
in items 4.1 through 4.26?

5

Critical
GLP

Sample Staining

5.1

Which stain to use and to follow manufacturer’s
instructions for FITC stain application?

14.2

14.2

5.3.2

Method Procedure

5.2

The slides are incubated in a humid chamber in the
dark at room temperature for approximately 30
minutes or per manufacturer’s directions?

14.3

14.3

5.3.3

Method Procedure

5.3

The working DAPI stain is prepared the day it is
used?

7.7.2

7.9.2

3.19.2

Method Procedure

5.4

The stock DAPI is stored at 1 to 10oC in the dark?

7.7.1

7.9.1

3.19.1

Method Procedure

5.5

The volume of working DAPI applied and the
incubation time?

14.6

14.6

-

Method Procedure

5.6

The technique used to drain the excess stain from
the well and to rinse the well?

14.5

14.5

-

Method Procedure

5.7

What type and amount of mounting media used?

7.8

7.10

-

Method Procedure

G-20

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
For each item, does the SOP specify:
5.8

5.9

Satisfactory
Classification

1623

1623.1

Cert

Yes

That all the edges of the cover slip are sealed well
with clear fingernail polish, unless Elvanol® is
used?

14.9

14.9

-

Method Procedure

The finished slides or slides not read immediately
are stored in a humid chamber in the dark at 1o to
10oC (humid chamber not required for Elvanol®)?

14.10

14.10

5.3.6

Method Procedure

5.10 Acceptable sample staining procedures, including
issues not noted in items 5.1 through 5.9?

Critical
GLP

6

Microscope and Examination

6.1

Instructions for ocular and Kohler adjustments?

10.3.4
10.3.6

10.7
10.8

3.22.10

Requirement

6.2

That all measurements must be recorded to the
nearest 0.5 micron?

15.2.2.3
15.2.3.3

15.2.2.4
15.2.3.4

3.22.5

Requirement

6.3

Microscope cleaning procedures?

10.4

10.9

3.22.11

Requirement

6.4

The recording of coordinates of all cysts and
oocysts on the worksheet for future reference; and
slide orientation on the microscope stage to
standardize coordinate recording?

-

-

-

Recommendation

The examination and acceptance of positive and
negative staining controls before proceeding with
examination of field samples?

15.2.1

15.2.1

5.4.6
5.4.7

Requirement

That each analyst characterizes 3 oocysts and 3
cysts on the positive staining control at each
examination session?

15.2.1.1

15.2.1.1

5.4.6

Requirement

-

-

5.4.8

Recommendation

15.2.2

15.2.2
15.2.3

5.4.9
5.4.10

Requirement

15.2

15.2.2.1
15.2.3.1

5.4.9.1
5.4.10.1

Requirement

6.5

6.6

6.7

Corrective actions if positive and/or negative
staining controls are not acceptable?

6.8

The criteria for organism identification?

6.9

Every positive organism in a field sample is
characterized and recorded?

6.10 Acceptable microscope and examination
procedures, including issues not noted in items 6.1
through 6.9?

7

Requirement
GLP

Reagents

G-21

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
For each item, does the SOP specify:
7.1

Procedures for the preparation of all essential
chemicals and reagents?

7.2

That expiration dates are specified for all reagents
prepared by the laboratory?

8

Satisfactory
Classification

1623

1623.1

Cert

Yes

7.0

7.0

4.2

Critical

-

-

4.2.2

Critical

Quality Assurance

8.1

Training protocol for new employees?

9.1

9.1

1.7

Requirement
GLP

8.2

Procedures for performing analyst verification?

10.6

9.10

7.1.9

Requirement
GLP

8.3

Positive and interfering organisms detected in field
samples are documented by photography?

-

-

5.4.11

Recommendation

8.4

Acceptable procedures for sample collection for
field or utility personnel?

-

-

6.1

Critical
GLP

8.5

Criteria for sample acceptance and corrective
action procedures?

8.1.3

8.1.3

6.

Requirement
GLP

8.6

Method required holding times?

8.2

8.2

6.4

8.7

Manual data recording procedures?

-

8.8

Procedures for checking the accuracy of data
transcriptions, including electronic data entry?

-

8.9

Procedures for checking the accuracy of manual
calculations?

-

-

Requirement
GLP

8.0

Critical
GLP

8.1

Critical
GLP

8.1

Critical
GLP

8.10 Procedures for electronic data entry and storage?

-

-

8.2

Critical
GLP

8.11 How backup of stored data is performed?

-

-

8.2

Critical
GLP

8.12 Corrective action procedures for OPR failures?

9.7.4

9.8.5

7.1.6.2

Requirement
GLP

8.13 Corrective action procedures for method blank
contamination?

9.6.2

9.7.3

7.1.5.2

Requirement
GLP

G-22

No

NA

UNK

Comments/
Response Requested

Reference*

Item to be Evaluated
For each item, does the SOP specify:

Satisfactory
Classification

1623

1623.1

Cert

Yes

8.14 Procedures for identifying and assessing declining
trends in recovery through review of control charts
and/or other recovery data?

-

-

7.1.7.2

Recommendation
GLP

8.15 Corrective action procedures for investigating QC
failures or declining trends in recovery?

-

-

7.1.7.2

Recommendation
GLP

8.16 Acceptable glassware washing procedures?

-

-

4.4

Critical
GLP

Comments:

G-23

No

NA

UNK

Comments/
Response Requested