PT Instructions

15 and 29 October 2018 Runs

PURPOSE

To provide for detailed instructions for participants in compliance with the requirements of ISO 17043, clause 4.6.1

INSTRUCTIONS

15 OCTOBER 2018 RUN

Scheme Description and Code 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 Storage
Antimicrobial susceptibility

(Mastitis)

JGKPT02

 2 x cultures ATB01M: Milk (liquid)

ATB02M: FD culture

 2-8oC ATB01M: Ready to process

ATB02M: Add 1 ml sterile D.H2O

 Test IMMEDIATELY upon receipt or no later than Wednesday 31 Oct. Culturing: Clin. micro

Susceptibility: Kirby-Bauer / Disc diffusion

 R.E.F 02

(supplied)

 Zone Ø in millimeters

Results expressed as:

S, I, R

 Culture: 1 rep

Susceptibility:

3 rep supplied discs;

1 rep own discs
18 x antibiotic discs Ticarcillin (x 6)

Pirlimycin (x 6)

Peni/Novobiocin (x 6)
Antimicrobial residues – meat

JGKPT24-26

 4 x pieces of chicken muscles

(>25 g)

 RES01F – RES04F 2-8oC Ready to process Test IMMEDIATELY upon receipt or no later than Wednesday 31 Oct. Rapid test

Microbiology

Chromatography

 R.E.F 24-26

(supplied)

 Presence/Absence of antimicrobials 1 rep
Antimicrobial residues – milk

JGKPT21-23

 4 x milk

(liquid)

 RES01M – RES04M 2-8oC Ready to process Test IMMEDIATELY upon receipt or no later than Wednesday 31 Oct. Rapid test

Microbiology

Chromatography

 R.E.F 21-23

(supplied)

 Presence/Absence of antimicrobials 1 rep
Food hygiene – meat (Detection)

JGKPT27-31

 3 x pieces of meat (chicken)

(>25 g)

 FH01 – FH03 2-8oC Ready to process Test IMMEDIATELY upon receipt or no later than Wednesday 31 Oct. Microbiology (conventional & automated) R.E.F 27-31

(supplied)

 Presence/Absence of target bacteria 1 rep
Mastitis pathogens – milk

JGKPT35

 2 x cultures MAST01 – FD culture

MAST02 – Milk (liquid)

 2-8oC ATB01M: Add 1 ml sterile D.H2O

ATB02M: Ready to process

 Test IMMEDIATELY upon receipt or no later than Wednesday 31 Oct. Culture & ID R.E.F 34-35

(supplied)

 Target mastitis bacteria 1 rep
Milk hygiene: TBC, Coliforms and E. coli count

JGKPT18-20

 5 x milk

(10 ml)

+

5 inoculums

 Samples ID:

MH01 – MH05

Inoculum ID:

MH inoculum 1 – 5

2-8oC See Instructions below Test IMMEDIATELY after mixing (see Instructions) 3M Petri Films R.E.F 18-20 For each samples test:

TBC

Coliforms

E. coli

 2 rep
Somatic cells counts – milk

JGKPT34

 6 x preserved milk

(20 ml)

 SCC01 – SCC06 2-8oC Mix thoroughly before testing Test within 5 days of dispatch Somatic cells counting device R.E.F 34-35

(supplied)

 Counts 3 rep
Mycotoxins – maize

JGKPT32 – 33

3 x maize meal

(100 g)

 MYC 01 – 03 2-8oC Ready to process Test within 5 days of dispatch ELISA

Chromatography

Other (specify)

 R.E.F 32-33

(supplied)

 Aflatoxin and Fumonisin

Presence / absence

Quantification

 Qualitative: 1

Quantitative: 1 (average)

29 OCTOBER 2018

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
Antimicrobial susceptibility

(Clinical)

JGKPT01

 2 x swabs

(+ 2  replicate bacterial suspensions)

 ATB01/C: Swine – Respiratory

ATB02/C: Dog – Otitis externa

 2-8oC Ready to process Test IMMEDIATELY upon receipt Disc diffusion technique R.E.F 1.2

(supplied)

 Zone Ø in millimeters.

Report reactions as: Sensitive (“S”), Intermediate (“I”), Resistance (“R”)

 Culture ID: 1 repl

Susceptibility:

JGK discs: 3 repl.

Lab discs: 1 repl.

(see result sheet)
Antimicrobial discs (x18) Florfenicol (x 6)

Clindamycin (x 6)

Oxytetracycline (x 6)

 Discs are ready to use (Commercial discs)
Brucella: RBT JGKPT03 6 FD bovine sera These samples are for Labs that perform RBT only

 

RBT01 – RBT06

 2-8oC Add 1 ml sterile DH2O per vial to samples 1, 2, 3, 5

Add 0.5 ml sterile DH2O per vial to samples 4, 6

Further instructions: see below

 Test IMMEDIATELY after reconstitution RBT only R.E.F 3.5

(supplied)

 Positive/Negative RBT 1 repl.
Brucella: RBT & CFT

 JGKPT03/04

 6 FD bovine sera These samples are for Labs that perform both RBT and CFT

 

RBT/CFT01 – RBT/CFT06

 2-8oC Add 1 ml sterile DH2O per vial to all samples

Further instructions: see below

 Test IMMEDIATELY after reconstitution RBT and CFT

(CFT: DAFF approved SOP for SA Labs)

 R.E.F 3.5

(supplied)

 RBT: Positive/Negative CFT: IU/ml RBT: 1 repl.

CFT: 2 repl.
Brucella: MRT JGKPT05 4 cow’s milk CAM01 – CAM04 2-8oC NA

Ready to process

 Preferably immediately but no later than Wednesday 31 Oct. MRT R.E.F 3.5

(supplied)

 Positive/Negative MRT 1 repl.
Brucella: Culture & Identification JGKPT06 2 milk (liquid)

1 x swab in Amies

 CAC01 & CAC02: Milk (liquid)

CAC03: Swab in Amies

 2-8oC NA

Ready to process

 Preferably immediately but no later than Friday 02 Nov. Conventional microbiology and

Molecular  technique

 R.E.F 06

(supplied)

 Presence/Absence: Brucella abortus

Strain: Field or vaccine

 1 repl.
Brucella: Slides

JGKPT07

 3 unstained smears Slide 1: x 3

Slide 2: x 3

Slide 3: x 3

 Room To

Dust-free

 Proceed with staining technique as per Lab SOP When and as convenient, but before submission deadline Stamp’s staining technique R.E.F 06

(supplied)

 Positive/Negative for Brucella 1 repl.
Trichomonas and Campylobacter ID by Culture JGKPT36-38 5 Cultures in transport media SW01 – SW05 2-8oC NA

Ready to process

 Preferably immediately but no later than Friday 02 Nov. Conventional microbiology R.E.F 36.9

(supplied)

 Presence/Absence: Trichomonas foetus, Campylobacter fetus 1 repl.
Trichomonas and Campylobacter ID by PCR

JGKPT37 & 39

 

 Molecular  technique Presence/Absence: Trichomonas foetus 1 repl.

Further Instructions

MILK HYGIENE:

Refrigerate milk samples and inoculums upon receipt

Before processing the samples, allow them to adjust to room temperature for 30 minutes. While samples are adjusting to room, reconstitute the inoculums by adding 1.5 ml sterile DH2O per vial.

Mix each vial well to break all lumps in the suspensions

Draw 50 µL from inoculum #1, add it to MH01 (i.e. sample 1)

Draw 50 µL from inoculum #2, add it to MH01 (i.e. sample 2)

Draw 50 µL from inoculum #3, add it to MH01 (i.e. sample 3)

Draw 50 µL from inoculum #4, add it to MH01 (i.e. sample 4)

Draw 50 µL from inoculum #5, add it to MH01 (i.e. sample 5)

Leave the inoculated milk at room temp for 5 minutes

Vortex the inoculated milk for 30 sec

Process the milk according to the Lab SOP

 

RBT ONLY (FOR LABS PERFORMING RBT ONLY):

Refrigerate samples upon arrival until used

Add 1 ml sterile D.H2O per vial to samples 1, 2, 3, 5. Mix well to break all lumps

Add 0.5 ml sterile D.H2O to samples 4, 6

Test each sample (RBT)

Mix sample 4 and sample 6 and use it as diluent to perform a two-fold dilution as follows:

Serially dilute (two-fold) sample 2 to extinction using the mixture of sample 4 and 6 as diluent

Test (RBT) all resulting serial samples until there is no agglutination (RBT-) and record results

 

RBT/CFT (FOR LABS PERFORMING BOTH RBT ABD CFT):

Refrigerate samples upon arrival until used

Add 1 ml sterile D.H2O per vial to all samples. Mix well to break all lumps

Test each sample (both RBT & CFT)

Mix sample 4 and sample 6 and use it as diluent to perform a two-fold dilution as follows:

Serially dilute (two-fold) sample 2 to extinction using the mixture of sample 4 and 6 as diluent

Test (both RBT & CFT) all resulting serial samples until the dilution where there is no agglutination (RBT-) and record results

 
Treatment of samples

No special treatment should be given to the PT samples. They [samples] should be handled in the same manner as all the other samples routinely tested in the laboratory.

 

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

All cultures must be handled in a biological safety cabinet.  Minimum requirement for JGKPT06 (Brucella Culture): Class 2+

 

Results recording

Only the supplied excel “Result entry forms” [R.E.F] should be used to capture the PT results. 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 or before the 19th of November 2018. 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