PT Instructions

WEEK 43: 21 & 22 OCTOBER 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

 

ATB01/C – Pig isolate

ATB02/C – Dog isolate

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

Remove seals and bungs from the vials and add 400 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 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:

Supplied discs: 3

(as per the result sheet)

Antimicrobial discs

(x 18+)

Cephalothin 30 µg; Oxoid  Lot # 3591986; exp. 03/12/2025.

Tilmicosin 15 µg; Oxoid Lot # 3751538; exp. 11/2026.

Amoxycillin/Clavulanic acid 30 µg; Oxoid Lot # 3714823; exp. 15/08/2024.

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: 500 µl

Sample/vial 2: 250 µl

Sample/vial 3: 250 µl

Sample/vial 4: 250 µl

Sample/vial 5: 500 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

 

 

 

 

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 FD cultures

 

CAC01 – CAC03 2-8oC Allow cultures and sDH2O to adjust to room temperature.

Remove seals and bungs from the vials and add 400 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 to facilitate discrete colonies.

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
JGKPT15

Bacteriology:

Pig Diseases

3 x FD cultures BAC01/P – BAC03/P 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 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 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
JGKPT16

Bacteriology: Sheep & Goat Diseases

3 x FD cultures BAC01/SG – BAC03/SG 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, 20th of November 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

 

FD: Freeze-Dried