PT Instructions
WEEK 31: 29 & 30 JULY 2024 RUN
PURPOSE
To provide for detailed instructions for participants in compliance with the requirements of ISO 17043, clause 4.6.1
INSTRUCTIONS
| Scheme
reference |
Samples | Samples Reconstitution and Techniques | Testing Timeframe | Approved Test Methods | Result Sheet for submission of Results to JGK | Unit of Measurement (Results) | Number of Test Replicates per sample | ||
| Type | Identification
(as per the labels) |
Storage | |||||||
| JGKPT01
Antimicrobial susceptibility (Clin. Pathogens) |
2 x FD cultures
Pus
2 x Amies swabs (BKP)
|
ATB01/C – cattle isolate
ATB02/C – cattle isolate |
2-8oC | Allow cultures and sDH2O to adjust to room temperature.
Remove seals and bungs from the vials and add 350 µl sDH2O to each vial. Allow the pellet to dissolve completely. Immediately, saturate a sterile swab with the reconstituted sample and inoculate 1/3 of the agar/culture plates (per the lab’s SOP). Streak with a sterile loop streak to facilitate discrete colonies.
Two ready-to-use Amies swabs are provided as BKP samples.
Proceed with Antibiogram testing as per the Lab’s SOP |
These samples are short-term samples; not intended for long-term storage.
Preferably, test the samples immediately upon receipt and reconstitution |
Disc diffusion technique
(Kirby Bauer) |
R.E.F 1.0
(available on www.jgklab.co.za) |
Zone Ø in millimeters.
Report results as: Sensitive (S), Intermediate (I), Resistance (R) |
Culture ID: 1
Susceptibility: Supplied discs: 3 (as per the result sheet) |
| Antimicrobial discs
(x 18) |
Compound sulphonamides 300 µg (x 7)/ Oxoid; Lot # 3233537; exp. 02/02/2024.
Kanamycin 30 µg (x 7)/ Oxoid; Lot # 3698274; exp. 12/07/2026. Apramycin 15 µg (x7)/ Oxoid; Lot # 3232842; exp. 03/2024. |
Discs are ready to use (Commercial discs) | |||||||
| JGKPT03
Brucella: RBT (Samples for Labs doing RBT only) |
6 x FD cattle sera | RBT01 – RBT06 | 2-8oC | Reconstitution:
Allow sera and sDH2O to adjust to room temperature. Remove seals and bungs, and add sDH2O as follows: Sample/vial 1: 250 µl Sample/vial 2: 250 µl Sample/vial 3: 250 µl Sample/vial 4: 500 µl Sample/vial 5: 250 µl Sample/vial 6: 250 µl Do not add cold water. |
Reconstituted samples must be mixed on the vortex and tested immediately.
There must be no clumps in the serum when tested |
RBT only | R.E.F 3-5
(available on www.jgklab.co.za) |
Positive (low, medium, high)
Or
Negative |
1 |
| JGKPT04
Brucella: RBT and CFT (Samples for Labs doing both RBT and CFT) |
6 x FD cattle sera | RBT/CFT01 – RBT/CFT06 | 2-8oC | Reconstitution:
Allow sera and sDH2O to adjust to room temperature. Remove seals and bungs and add 500 µl sDH2O to each sample/vial.
Do not add cold water. |
Reconstituted samples must be mixed on the vortex and tested immediately.
There must be no clumps in the serum when tested |
RBT and CFT | R.E.F 3-5
(available on www.jgklab.co.za) |
RBT:
Positive (low, medium, high) Or Negative |
1 |
| CFT:
Option 1: Endpoint reaction (e.g. 1/4 ++++, 1/128+)
Option 2: Titer (e.g. 24, 480)
NB: the corresponding serum dilution (e.g. 1/4) must always be indicated or ticked on the result sheet |
2
|
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| JGKPT05
Brucella: MRT |
4 x bulk tank milk
(not preserved) |
CAM01 – CAM04 | 2-8oC | N/A
Ready to process |
Test immediately upon receipt | MRT | R.E.F 3-5
(available on www.jgklab.co.za) |
Positive (weak, strong) Or
Negative |
1 |
| JGKPT06
Brucella: Culture & Identification |
3 x cow’s milk
1 x Amies swab
|
CAC01 – CAC04 | 2-8oC | N/A
Ready to use |
These samples are short-term samples; not intended for long storage.
Preferably, test the samples immediately upon receipt. |
Conventional microbiology
and
Molecular technique |
R.E.F 06
(available on www.jgklab.co.za) |
Presence or Absence of Brucella abortus
Strain: Field or vaccine |
1 |
| JGKPT07
Brucella Slides |
6 x unstained smears | BA Slide 01: x 2
BA Slide 02: x 2 BA Slide 03: x 2 |
Room To
Dust-free |
Proceed with staining technique as per the laboratory’s SOP | Test immediately upon receipt | Stamp’s staining technique | R.E.F 06
(available on www.jgklab.co.za) |
Positive or Negative for Brucella | 1 |
| 3 x FD cultures | LM01 – LM03 | 2-8oC | Reconstitution:
Allow cultures and sDH2O to adjust to room temperature. Remove seals and bungs and add 400 µl sDH2O to each Sample/vial.
Allow the pellet to dissolve completely. Immediately, saturate a sterile swab with the reconstituted sample and inoculate 1/3 of the agar/culture plates (per the lab’s SOP). Streak with a sterile loop streak to facilitate discrete colonies. |
These samples are short-term samples; not intended for long storage.
Preferably, test the samples immediately upon receipt and reconstitution |
Conventional microbiology
and
Molecular technique |
R.E.F 08-10
(available on www.jgklab.co.za) |
Presence or absence of Listeria.
Listeria: Full identification of Listeria (Genus and species)
Listeria: Identify Genus and species (e.g. Listeria monocytogenes) |
1 | |
| JGKPT09
Bacteriology: Poultry Salmonella (Clinical Micro) |
3 x FD cultures | SAP01 – SAP03 | 2-8oC | Reconstitution:
Allow cultures and sDH2O to adjust to room temperature. Remove seals and bungs and add 400 µl sDH2O to each Sample/vial.
Allow the pellet to dissolve completely. Immediately, saturate a sterile swab with the reconstituted sample and inoculate 1/3 of the agar/culture plates (per the lab’s SOP). Streak with a sterile loop streak to facilitate discrete colonies. |
These samples are short-term samples; not intended for long storage.
Preferably, test the samples immediately upon receipt and reconstitution |
Conventional microbiology
and
Molecular technique |
R.E.F 08-10
(available on www.jgklab.co.za) |
Presence or absence of Salmonella.
Salmonella: Full identification of Salmonella (Genus and serotype) |
1 |
| JGKPT10
Bacteriology: Mammals Salmonella (Clinical Micro) |
3 x FD cultures | SAM01 – SAM03 | 2-8oC | 1 | |||||
| JGKPT35 & 37
Trichomonas and Campylobacter ID by Culture |
5 x Cultures in transport media | SW01 – SW05 | 2-8oC | N/A
Ready to process |
Test immediately upon receipt | Conventional microbiology | R.E.F 35-38
(available on www.jgklab.co.za) |
Presence or Absence of
Trichomonas foetus/ Campylobacter fetus |
1 |
| JGKPT36 & 38
Trichomonas and Campylobacter ID by PCR |
Molecular technique | 1 | |||||||
|
Treatment of samples No special treatment should be given to the PT samples. They [PT samples] should be handled in the same manner as all the other samples routinely tested in the laboratory, including TAT
Handling and safety requirements Contact surfaces must be decontaminated with 70% alcohol or other suitable disinfectants. Used laboratory utensils must be cleaned and sterilized as per laboratory standard operating procedure. Unused materials and waste must be removed as per laboratory waste management protocol. This involves autoclaving and/or removal by an approved waste removal company.
Specific environmental conditions required for the participant to conduct tests JGKPT06 (Brucella Culture) must be handled in a biological safety cabinet class 2+
Results recording Only the supplied Excel “Result entry forms” [R.E.F] should be used to capture the PT results. The R.E.F grouped into WK8,19,31,43 and WK9,20,32,44 are available online (www.jgklab.co.za) and can be downloaded under “PUBLICATIONS” The Participants must fill in the cover page and answer all questions before capturing the results on R.E.F.
Submission of PT results to JGK All results must be submitted on Wednesday, 21st of August 2024. Any results received after this date will not be included in the analysis.
Enquiries For any inquiry, please use the contact details provided above to contact JGK
Return of the proficiency test items None
|
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FD: Freeze-Dried