Milk Safety References
National Conference on Interstate Milk Shipments (NCIMS)

APPENDIX N BULK MILK TANKER SCREENING TEST FORM
Charm SL (raw bovine and caprine) and SL-6 (raw bovine)
Beta-Lactam Tests
GENERAL REQUIREMENTS

1. __________ See Appendix N General Requirements form items 1-8 & 15

   SAMPLES

2. __________ See Appendix N General Requirements (GR) form item 9

   APPARATUS & REAGENTS

3. __________ Equipment
   1. __________ Strip Incubator, 56±1 0 C SL Beta-lactam test; 45±2 0 C SL–6 Beta-lactam Test
      1. __________ Level. Temperature checked daily (day of use), records maintained
      2. __________ thermometer, for each incubator (App N GR #3)
      3. __________ Lid closed (spring ajar), when not running tests
      4. __________ Incubator Temperature: ______________
   2. __________ ROSA Reader or equivalent with print out or download of data. ROSA Reader V1.07
      or higher for SL–6, or equivalent capable of reading
      3 line test, Manual Available Serial Number: ______________
      1. __________ Calibrators – 2 for each assay– 2 lines for SLBL and/or 3 lines for SL–6.
         

         Range(s)
         Two Line:
         Low:_________________________
         High_________________________
         
         Three Line:
         Low:_________________________
         High:________________________

      2. __________ Calibrator serial numbers match ROSA reader SN
      3. __________ Do not proceed if out of range. Manufacturer should be contacted for corrective actions
      4. __________ Printer or computer link for hardcopy download
      5. __________ Records maintained
   3. __________ Pipettor- 300 µL and disposable tips, calibrated (see App. N GR item 7)
   4. __________ Or s ingle use 300 µL poly-pipet with overflow bulb to accurately measure amount of sample, supplied by manufacturer (screening only)
   5. __________ Optional Timer – if not built into Incubator
   6. __________ Optional Centrifuge (Not applicable to SL–6 Beta-lactam Test)- mini or equivalent (1200– 2000 x g) for frozen controls
4. __________ Reagents
   1. __________ Test Strips

      __________Lot #: _______ Exp. Date: _______

      __________Rec'd: _______ QC Date: _______by _______

   2. __________ Positive Control
      1. __________ Lyophilized or tablet 5 ppb Penicillin G for SL Beta–lactam Test

         __________Lot #: _______ Exp Date: _______ ________

      2. __________ Lyophilized or tablet 5ppb Penicillin G/10ppb cloxacillin for SL–6 Beta-lactam Test

         __________Lot # _______ Exp. Date:_______ ________

      3. __________ Do not proceed if out of range. Manufacturer should be contacted for corrective actions
      4. __________ Printer or computer link for hardcopy download
      5. __________ Records maintained
   3. __________ Negative Control
      1. __________ Previously negative tested raw milk (item 5b)
5. __________ Reagent stability
   1. __________ SL–6 reagents received refrigerated.
   2. __________ Reagents stored at 0–4.4 0 C, maintain no longer than manufacturer's expiration date
   3. __________ Negative Control – raw milk tested –600 or more negative with SL Beta-lactam Test or –400 or morenegative with SL–6 Beta–lactam

      __________Test Value:________ Do not proceed if out of range.

      1. __________ Used within 72 hours when maintained at 0–4.4 0 C
      2. __________ Test for suitability each time, records maintained
      3. __________ Or, frozen immediately (within 6 hours) and stored in a non-frost-free freezer, or in a styrofoam container in a frost free freezer, for no more than 2 months at –15 0 C or below Lab

         __________Date prep:._______ Lab Exp. Date: _______ ________

         1. __________ For SLBL ONLY, not applicable to SL-6 thaw, centrifuge 3 minutes, cool and use within 24 hours. Do not refreeze.
   4. __________ Positive Control- Manufacturer supplied, maintain no longer than manufacturer's expiration date
      1. __________ Reconstituted with Negative Control (raw milk), used within 48 hours when maintained at 0–4.4 0 C
      2. __________ Test for suitability each time prepared and/or day of use, must produce +400 or greater reading, records maintained.

         __________Test Value:________ Do not proceed if out of range. ________

      3. __________ Or, frozen immediately (within 6 hours) and stored in a non-frost-free freezer, or in a styrofoam container in a frost-free freezer, for no more than 2 months at –15 0 C or below

         __________Lab Date prep: _______ Lab Exp. Date: _______ ________

         1. __________ Thaw, centrifuge 3 minutes( not applicable to SL–6 Beta-lactam Test), cool and use within 24 hours. Do not refreeze.

    

   TECHNIQUE

6. __________ Daily Performance and Operation Check
   1. __________ See App. N GR item 10)
   2. __________ Reader Versions 1.5 and higher, use Escape 5 reader function to enter performance monitor mode of reader- reference manual for directions
   3. __________ Check Calibrators, records maintained
   4. __________ Controls give appropriate readings in performance monitoring mode (reader version 1.05 and higher) prior to any sample analysis. If out of range, manufacturer should be contacted for corrective action, 800-343-2170.
   5. __________ Do not proceed if out of range.
7. __________ Reagent stability
   1. __________ Label Test Strips, one for each test sample and each control
   2. __________ Place strip into appropriate incubator, avoid crushing sample compartment. For multiple samples,complete steps b-f for each sample/control, before starting test of next sample.
   3. __________ While holding strip flat, peel (to 'peel to here' line) back plastic to expose sample pad compartment
   4. __________ Mix milk sample or control 25 times in seven seconds with a 1 ft movement or vortex control for 10 seconds when appropriate
   5. __________ Slowly pipette 300 µL sample to either side of the sample pad compartment(if applicable, use etched line on incubator as location guide). Avoid contact of sponge with pipet tip.
      1. __________ Draw up sample, avoiding foam and bubbles.(with 300ul calibrated pipet only pre-wet tip by expelling sample and drawing up again)
      2. __________ Or for Screening only use poly pipet
         1. __________ While holding poly pipet almost vertically with bulb side pointing down, depress topbulb and insert into sample (below foam).
         2. __________ Release bulb and sample should completelyfill pipet shaft and overflow into bottom ½ of lower bulb
         3. __________ Pipet sample in pipet shaft by depressing top bulb only into test strip while leaving excess sample in overflow bulb
   6. __________ Re-seal plastic firmly around sample pad compartment
   7. __________ Repeat Step 7b-f for each sample and/or control test strip in incubator
   8. __________ Close and latch incubator cover. If no automatic timer in incubator, set timer for 8 minutes, do not exceed 10 minutes
   9. __________ At end of incubation, visually inspect C (Control) line, an absent C line or a partial C line or an indistinct C line indicates an invalid test and the sample/control must be re–tested
   10. __________ Insert only valid test(s) in ROSA reader (set to appropriate channel.
       1. __________ SLBL slow blink for SL Beta–lactam Test
       2. __________ SLBL rapid Blink for SL–6 Beta–lactam Test
   11. __________ Press ENTER, reading and interpretation appear in 5 seconds, read strips within 10 minutes of completion of incubation
8. __________ Interpretation with ROSA Reader
   1. __________ If there is a negative or zero reading on the reader, sample is Negative (NF)
   2. __________ If there is a positive reading on the reader, sample is an Initial Positive
9. __________ Verification of Initial Positive Samples (see App. N GR item 11); Confirmation of PresumptivePositive Samples (see App. N GR item 12); and Producer Traceback (see App. N GR item 13)
10. __________ Reporting (see App. N GR item 14) .
    1. __________ Reader tapes or computer print outs maintained