PT Instructions

WEEK 19: 08 & 09 MAY 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 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 Grown Bacteria on NA Plates (main samples)

2 x Cultures on Amies swabs (BKP samples)

NB: the bacteria on NA plates and on the swabs are identical

ATB01/C – cattle

ATB02/C – chickens

2-8oC ATB01/C: Ready to use

ATB02/C: Ready to use

Test the samples immediately upon receipt 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) Cephazolin 30 µg (x 7)/ Oxoid; Lot # 3195880; Exp. 09/12/2023

Gentamycin 10 µg (x 7)/ Oxoid; Lot # 3108138; Exp. 30/07/2023

Tetracycline 30 µg (x 7)/ Oxoid; Lot # 3214586; Exp. 14/01/2024

Discs are ready to use (Commercial discs)
JGKPT03

Brucella: RBT

(Samples for labs doing RBT only)

6 FD cattle sera RBT01 – RBT06 2-8oC Samples 1, 2, 4 and 5:

Add 500 µl sDH2O to the vial

 

Sample 3:

Add 1 ml sDH2O to the  vial

Sample 6:

Add 250 µl sDH2O to the  vial

Once reconstituted, samples must be tested without delay 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 FD cattle sera RBT/CFT01 – RBT/CFT06 2-8oC Sample 1:

Add 2 ml sDH2O to the vial

 

Samples 2, 4, and 5:

Add 500 µl sDH2O to the vial

 

Sample 3:

Add 1 ml sDH2O to the vial

 

Sample 6:

Add 700 µl sDH2O to the  vial

Once reconstituted, samples must be tested without delay 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 bulk tank milk

(not preserved)

CAM01 – CAM04 2-8oC NA

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

CAC01:

Amies swab

CAC02:

FD culture

CAC03:

Bovine milk

CAC04:

Amies swab

CAC01 – CAC04 2-8oC FD cultures:

Add 1 ml sD.H2O to each vial

 

Swabs:

Ready to process

 

Milk:

Ready to process

Once reconstituted, sample must be tested without delay 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 unstained smears Slide 1: x 2

Slide 2: x 2

Slide 3: x 2

Room To

Dust-free

Proceed with staining technique as per Lab SOP Test immediately upon receipt Stamp’s staining technique R.E.F 06

(available on www.jgklab.co.za)

Positive/Negative for Brucella 1
JGKPT11

Bacteriology: Diseases of cattle

Sample 1 (BA01/C):

Grown Bacteria on NA Plate (main sample) + Amies swab (BKP)

Sample 2 (BA02/C):

Grown Bacteria on NA Plate (main sample) + Amies swab (BKP)

Sample 3 (BA03/C):

Grown Bacteria on NA Plate (main sample) + Amies swab (BKP)

Sample 4 (BA04/C):

Bovine quarter milk

 

NB: the bacteria on NA plates and on the swabs are identical

BA01/C – BA04/C

Disease scenarios available on a separate document

2-8oC NA plates:

Ready to process

 

Amies swabs:

Ready to process

 

Milk:

Ready to process

 

Test immediately upon receipt Conventional microbiology and

Molecular  technique

R.E.F 11-16

(available on www.jgklab.co.za)

Full bacterial identification 1
JGKPT12

Bacteriology: Diseases of chickens

Sample 1 (BA01/CH):

Grown Bacteria on NA Plate (main sample) + Amies swab (BKP)

Sample 2 (BA02/CH):

Grown Bacteria on NA Plate (main sample) + Amies swab (BKP)

Sample 3 (BA03/CH):

Grown Bacteria on NA Plate (main sample) + Amies swab (BKP)

NB: the bacteria on NA plates and on the swabs are identical

BA01/CH – BA03/CH

Disease scenarios available on a separate document

2-8oC NA plates:

Ready to process

 

Amies swabs (BKP):

Ready to process if necessary

 

Test immediately upon receipt 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 Cultures in transport media SW01 – SW05 2-8oC 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, 31st of May 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