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sops > E-0003 - Presence/Absence (PA) > Flashcards

E-0003 - Presence/Absence (PA) Flashcards

1
Q

What is the code for P/A?

A

SM 9223-B

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2
Q

What is the first thing you do?

A

Make sure I have proper PPE on
- Lab coat, goggles, and gloves

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3
Q

What do you do after putting on PPE?

A

Prepare the logbook

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4
Q

Preparing the logbook: what information do you include?

A
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5
Q

What do you do after preparing the logbook?

A

Sterilize the work bench area, equipment, and gloves with 70% IPA

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6
Q

Describe the setup for a PA.

A
  1. In rack:
    - 3 QC bottles at the top: EC, KP, and PA
    - Samples along the bottom, left to right, 5 in a row
  2. Paper towels
    - Lay media packs on them, plastic side up
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7
Q

What do you do after preparing the logbook?

A

Sterilize the work bench area, equipment, and gloves with 70% IPA

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8
Q

What do you do after sterilizing work area/equipment/gloves?

A

Check sample volume (should be 100 mL)
- If exceeds: pipette off excess
- If less than 95 mL: tell PM immediately
- If overfilled: transfer to sterile container to homogenize and transfer back to sample bottle

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9
Q

What must you do before pipetting off excess sample volume?

A

Shake the sample 25 times

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10
Q

What do you do after checking sample volumes?

A

Acquire the appropriate media

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11
Q

When do you use Colilert 18 vs. Colilert 24 vs. Colisure?

A
  • Colilert 18: after 2pm
  • Colilert 24: before 2pm
  • Colisure: if sample has tint (put in different batch, has its own QCs)
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12
Q

What do you do after acquiring the appropriate media?

A
  • Check residual chlorine of samples that arrived without having had residual chlorine checked in the field
  • If no chlorine check, add media (snap open packets and pour in, don’t shake)
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13
Q

How do you check residual chlorine if needed?

A
  • Shake sample 25 times before opening
  • Use sterile transfer pipette to drop sample onto chlorine test strip
  • If deep blue: chlorine present
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14
Q

What should you do with the media before adding it to the samples?

A

Spray the front of packet (plastic side) with 70% IPA

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15
Q

What are the protocols for adding media?

A
  • Snap open packet (hold away from you)
  • Keep 1 inch distance when pouring
  • Don’t shake packet
  • Recap sample and gently shake 10 times to dissolve media
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16
Q

What do you after adding the media?

A

Place sample back in the rack in the order you took them out

17
Q

What do you do after putting the samples back in the rack after adding the media?

A

Prepare the QCs

18
Q

What do you do after preparing the QCs?

A

Put a batch sticker on top of the EC QC bottle
- On batch sticker: day of week (abbreviation), batch setup alphanumeric letter for the day, analysis
- Ex) Mon, A, P/A
- Put a matching batch sticker in the logbook (top right corner)

19
Q

What do you after adding the batch sticker?

A
  • Colilert 18: place in water bath at 44.5 +/- 0.2 degrees C for 7 minutes (record water bath in time and temp and water bath ID)
  • Colilert 24 and Colisure: incubate at 35 +/- 0.5 degrees C for 24-28 hours (record incubator in time and temp and incubator ID)
20
Q

At what temp and for how long is Colilert 18 in the water bath and incubator?

A
  1. Water bath
    - 44.5 +/- 0.2°C
    - 7-10 min
  2. Incubator
    - 35 +/- 0.5°C
    - 18-22 hours
21
Q

At what temp and for how long are Colilert 24 and Colisure in the incubator?

A
  • 35 +/- 0.5°C
  • 24-28 hours
  • Colisure can be up to 48 hours if needed
22
Q

What do you do after putting the samples in the incubator?

A

Calculate the reading time based off the time you put it into the incubator (or water bath for Colilert 18)
- Write on post-it and put in the middle of the page in the logbook

23
Q

How do you set up the QCs?

A
  1. Inoculate with the appropriate bacteria (EC, KP, and PA)
  2. Pour sterile RGW into sterile 120 mL bacti bottle
  3. Use a loop, dip into an already prepared culture, transfer the loop to the bottle and stir back and forth, discard loop in biohazard bag, cap bottle
  4. Add media packet
24
Q

Preparing the logbook for reading: what information do you include?

A
25
Q

After setting up the logbook for reading, what do you do?

A

Read the QCs first and record the results
- EC: color change and fluorescence
- KP: color change but not fluorescence
- PA: no color change and no fluorescence

26
Q

What do you do after reading the QCs?

A

Read the samples and record the results
- Check for color change: + or - under “Total Coliforms”
- Check for fluorescence: + or - under “E. coli”

27
Q

What do you do after reading the samples and recording the results?

A
  • Cross out the post-it and move it to the bottom of the page
  • Cross out the to-do list
28
Q

What do you do after crossing out the post-it and to-do list after reading?

A

Put the samples and QCs on the 24-hour hold shelf

29
Q

List the general steps for a PA (not including reading).

A
  1. PPE
  2. Get samples
  3. Prep logbook
  4. Sterilize work area
  5. Check sample volume
  6. Sterilize media packets and add media
  7. Prep QCs
  8. Batch sticker
  9. Incubate
  10. Calculate reading time
  11. Sterilize work area

sops (8 decks)

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