PT Instructions

WEEK 43: 23 OCTOBER & 24 OCTOBER 2023 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 Sheet to be used 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

1 x NA Plate with 2 grown bacteria

Notes:

1. Culturing must be done from the FD cultures (see reconstitution)

2. The NA plate can be used as BKP but its main purpose is to serve as QC evidence for the growth of the FD cultures

ATB01/C – Sheep

ATB02/C – Pig

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 (plates as per lab’s SOP). Using a sterile loop streak to facilitate discrete colonies

These samples are not intended to be stored.

Preferably, test the samples immediately upon receipt and upon 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 result sheet)

Antimicrobial discs

(x 21)

Sulfonamides 300 µg (x 7)/ Oxoid; Lot # 3233537; Exp. 02/02/2024

Apramycin 15 µg (x 7)/ Oxoid; Lot # 3232842; Exp. 03/2024

Cephazolin 30 µg (x 7)/ Oxoid; Lot # 3195880; Exp. 09/12/2023

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: 300 µl

Sample/vial 2: 250 µl

Sample/vial 3: 350 µl

Sample/vial 4: 250 µl

Sample/vial 5: 1 ml

Sample/vial 6: 500 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/Negative 1
JGKPT04

Brucella: RBT/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 sDH2O as follows:

Sample/vial 1: 500 µl

Sample/vial 2: 500 µl

Sample/vial 3: 500 µl

Sample/vial 4: 500 µl

Sample/vial 5: 300 µl

Sample/vial 6: 500 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 and CFT R.E.F 3-5

(available on www.jgklab.co.za)

RBT: Positive/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/Negative 1
JGKPT06

Brucella: Culture & Identification

3 x FD cultures

 

CAC01 – CAC03 2-8oC Reconstitution:

Allow cultures and sDH2O to adjust to room temperature.

Remove seals and bungs, and add sDH2O as follows:

Sample/vial 1: 400 µl

Sample/vial 2: 400 µl

Sample/vial 3: 400 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 (plates as per lab’s SOP). Using a sterile loop streak to facilitate discrete colonies

These samples are not intended to be stored.

Preferably, test the samples immediately upon receipt and upon reconstitution

Conventional microbiology

 

and

 

Molecular  technique

R.E.F 06

(available on www.jgklab.co.za)

Presence/Absence: 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/Negative for Brucella 1
JGKPT13

Bacteriology: Diseases of Pig

3 x FD cultures

1 x NA Plate with 3 grown bacteria

Notes:

1. Culturing must be done from the FD cultures (see reconstitution)

2. The NA plate can be used as BKP but its main purpose is to serve as QC evidence for the growth of the FD cultures

BA01/P – BA03/P

Disease scenarios available on a separate document

2-8oC Reconstitution:

Allow cultures and sDH2O to adjust to room temperature.

Remove seals and bungs, and add sDH2O as follows:

Sample/vial 1: 400 µl

Sample/vial 2: 400 µl

Sample/vial 3: 400 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 (plates as per lab’s SOP). Using a sterile loop streak to facilitate discrete colonies

These samples are not intended to be stored.

Preferably, test the samples immediately upon receipt and upon reconstitution

Conventional microbiology

 

and

 

Molecular  technique

R.E.F 11-16

(available on www.jgklab.co.za)

Full bacterial identification 1
JGKPT13

Bacteriology: Diseases of Sheep & Goats

3 x FD cultures

1 x NA Plate with 3 grown bacteria

Notes:

1. Culturing must be done from the FD cultures (see reconstitution)

2. The NA plate can be used as BKP but its main purpose is to serve as QC evidence for the growth of the FD cultures

BA01/SG – BA03/SG

Disease scenarios available on a separate document

2-8oC Reconstitution:

Allow cultures and sDH2O to adjust to room temperature.

Remove seals and bungs, and add sDH2O as follows:

Sample/vial 1: 400 µl

Sample/vial 2: 400 µl

Sample/vial 3: 400 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 (plates as per lab’s SOP). Using a sterile loop streak to facilitate discrete colonies

These samples are not intended to be stored.

Preferably, test the samples immediately upon receipt and upon reconstitution

Conventional microbiology

 

and

 

Molecular  technique

R.E.F 11-16

(available on www.jgklab.co.za)

Full bacterial identification 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/Absence:

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, 15th of November 2023. 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