PT Instructions

WEEK 31: 28 & 29 JULY 2025 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

2 x Amies transport swabs (BKP samples)

 

ATB 01/C – Dog isolate

ATB 02/C – Dog isolate

2-8oC Allow cultures and sDH2O to adjust to room temperature.

Remove seals and bungs from the vials and rehydrate the cultures with sDH2O as follows: ATB01: 650 µl; ATB02: 450 µl.

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 to facilitate discrete colonies.

 

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:

Discs supplied: 3

(as per the result sheet)

Antimicrobial discs

(x 18)

Oxytetracycline 30 µg; Oxoid 6265879; exp. 14/05/2028.

Sulphamethoxazole/Trimethoprim 25 µg; Oxoid 3552294; exp. 15/09/2025.

Clindamycin 2 µg; Oxoid 6211907; exp. 13/01/2028.

Discs are ready to use (Commercial discs)
JGKPT03

Brucella: RBT

(Samples for Labs doing RBT only)

6 x FD cattle sera RBT 01 – RBT 06 2-8oC Reconstitution:

Allow sera and sDH2O to adjust to room temperature.

Remove seals and bungs, and add sDH2O as follows:

Sample/vial 1: 400 µl

Sample/vial 2: 500 µl

Sample/vial 3: 400 µl

Sample/vial 4: 400 µl

Sample/vial 5: 400 µl

Sample/vial 6: 500 µl

 

Do not add cold water.

Reconstituted samples must be mixed on a 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/CFT 01 – RBT/CFT 06 2-8oC Reconstitution:

Allow sera and sDH2O to adjust to room temperature.

Remove seals and bungs, and add sDH2O as follows:

Sample/vial 1: 400 µl

Sample/vial 2: 500 µl

Sample/vial 3: 400 µl

Sample/vial 4: 400 µl

Sample/vial 5: 400 µl

Sample/vial 6: 500 µl

 

Do not add cold water.

Reconstituted samples must be mixed on a 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., 15, 581)

NB: the corresponding serum dilution (e.g. 1/4) must always be indicated or ticked on the result sheet

 

 

 

2

 

 

 

 

JGKPT05

Brucella: MRT

4 x bulk tank milk

(not preserved)

CAM 01 – CAM 04 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 cows’ milk

 

CAC 01 – CAC 03 2-8oC N/A

Ready to process

These samples are short-term samples, not intended for long storage.

Please 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 the 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
JGKPT09

Clinical Microbiology:

Salmonella - Poultry

3 x FD cultures

3 x Amies transport swabs (BKP samples)

SAP 01 – SAP 03 2-8oC FD cultures:

Allow cultures and sDH2O to adjust to room temperature.

Remove seals and bungs and rehydrate the cultures by adding sDH2O to the vials as follows:

 

SAP 01:  650 µl

SAP 02:  450 µl

SAP 03:  650 µl

 

SAM 01:  650 µl

SAM 02:  450 µl

SAM 03:  450 µl

 

BA 01/DC: 450 µl

BA 02/DC: 650 µl

BA 03/DC: 650 µl

 

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 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 09-10

(available on www.jgklab.co.za)

Presence or absence of salmonella and full identification of the salmonella isolates 1
JGKPT10

Clinical Microbiology:

Salmonella – Mammals

3 x FD cultures

3 x Amies transport swabs (BKP samples)

SAM 01 – SAM 03 2-8oC 1
JGKPT13

Bacteriology:

Dog & Cat Diseases

3 x FD cultures

3 x Amies transport swabs (BKP samples)

BA 01/DC – BA 03/DC 2-8oC R.E.F 11-16

(available on www.jgklab.co.za)

Full bacterial identification of the isolates and indication of the bacteria responsible for the conditions described in the scenarios 1
JGKPT14

Bacteriology: Horse Diseases

Will be shipped on WK32
JGKPT35 & 37

Trichomonas and Campylobacter ID by Culture

5 x broths SW 01 – SW 05 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

Campylobacter fetus

1
JGKPT36 & 38

Trichomonas and Campylobacter ID by PCR

5 x broths SW 01 – SW 05 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 the laboratory standard operating procedure. Unused materials and waste must be removed as per the 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, 20th of August 2025. 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

 

FD: Freeze-Dried