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