PT Instructions

WEEK 44: 30 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 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

(as per the labels)

 Storage
Antibiogram – mastitis pathogens

JGKPT02

 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/M – Dairy cow

ATB02/M – Dairy cow

 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

 Culturing: Micro

Susceptibility: Disc diffusion (Kirby-Bauer) / Qualitative

 

 

 R.E.F 02

(supplied)

 Zone Ø in millimeters.

Report results as:

Sensitive (S), Intermediate (I), Resistance (R)

 Culture: 1 rep

Susceptibility:

3 replicates (supplied discs)
Antimicrobial discs

(x 21)

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

Florfenicol 30 µg (x 7)/ Oxoid; Lot # 3274026; Exp. 04/2022

Penicillin G 10 IU (x 7)/ Oxoid; Lot # 3164322; Exp. 10/2023

 Commercial discs:

Ready to use
Antimicrobial residues – meat

JGKPT23-25

 4 x Fresh samples

(chicken liver)

 RES-01-F – RES-04-F 2-8oC Fresh samples: Ready to process Test immediately upon receipt Rapid test

Microbiology

Chromatography

 R.E.F 23-25

(supplied)

 Presence/Absence of antimicrobials

 

ID of the antibiotic

 1 rep
Antimicrobial residues – milk

JGKPT20-22

 4 x bovine milk RES-01-M – RES-04-M 2-8oC N/A

Ready to process

 Test immediately upon receipt Rapid test

Microbiology

Chromatography

 R.E.F 20-22

(supplied)

 Presence/Absence of antimicrobials

 

Name of the antibiotic

 1 rep
Food hygiene – Detection

JGKPT25-30

 4 x Amies swabs

1 x NA plate with 4 grown bacteria

 FH01 – FH04 2-8oC N/A

Ready to process

 Test immediately upon receipt Microbiology (conventional & PCR) R.E.F 25-30

(supplied)

 Presence/Absence of target bacteria 1 rep
Food hygiene – Enumeration 4 x FD samples FH01/Counts – FH04/Counts 2-8oC Allow samples to adjust to room temperature. Add 6 ml preheated (37oC) sterile DH2O to each jar. Mix content thoroughly on a vortex until there is no clumps Test immediately upon receipt/

reconstitution

 Counts TBC: 2 rep

Coliforms: 2 rep

E. coli: 2 rep
Mastitis pathogens – milk

JGKPT34

 2 x mastitis FD cultures MAST01 – MAST02 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

 Test immediately upon receipt/

reconstitution

 Culture & ID R.E.F 33-34

(supplied)

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

JGKPT17-19

 4 x FD milk (bovine) MH01 – MH04 2-8oC Add 6 ml sterile DH2O

to each vial

Notes:

Do not use cold water. Samples must adjust to room temperature for ≥ 30 min before adding sD.H2O.

D.H2O must be at 37oC in a water bath for ≥30 min before use. Mix content thoroughly on a vortex until there is no clumps.

 Test immediately upon receipt/

reconstitution

 3M Petri Films

Conventional technique

 R.E.F 17-19 Counts TBC: 2 rep

Coliforms: 2 rep

E. coli: 2 rep
Aflatoxins & Fumonisins

JGKPT31-32

 3 x maize meal MYC01 – MYC03 2-8oC N/A

Ready to process

 Test immediately upon receipt Chromatography R.E.F 31-32 Detection & quantification of Aflatoxins (ppb) and Fumonisins (ppm) 1 rep.
BVD & Fasciola

JGKPT39-40

 5 x FD sera (bovine) BVD/FAS01 – BVD/FAS05 2-8oC Allow samples and sDH2O to adjust to room temperature. Add s DH2O to the vials as follows:

Sample 1: 300 µL

Sample 2: 300 µL

Sample 3: 300 µL

Sample 4: 350 µL

Sample 5: 300 µL

 Test immediately upon receipt/

reconstitution

 ELISA R.E.F 39-45 Elisa ODs 2 rep.
Avian influenza - Abs

JGKPT41

 6 x FD sera (chicken) AI01 – AI06 2-8oC Allow samples and sDH2O to adjust to room temperature. Add sDH2O to the vials as follows:

Sample 1: 500 µL

Sample 2: 500 µL

Sample 3: 350 µL for labs doing one test (ELISA or HI)

Sample 3: 700 µL for labs doing both ELISA & HI; supplied as 1 x 700 µL (written on the vial) or 2 x 350 µL (to be mixed)

Sample 4: 500 µL

Sample 5: 500 µL

Sample 6: 500 µL

 Test immediately upon receipt/

reconstitution

 ELISA

HA/HI

 R.E.F 39-45 ODs

Titres

 2 rep
Infectious bronchitis virus – Abs

  JGKPT42

 6 x FD sera (chicken) IBV01 – IBV06 2-8oC Allow samples and sDH2O to adjust to room temperature. Add sDH2O to the vials as follows:

Sample 1: 200 µL

Sample 2: 200 µL

Sample 3: 200 µL

Sample 4: 200 µL

Sample 5: 200 µL

Sample 6: 200 µL

 Test immediately upon receipt/

reconstitution

 ELISA

HA/HI

 R.E.F 39-45 ODs

Titres

 2 rep
Newcastle disease – Abs JGKPT43 6 x FD sera (poultry) NDV01 – NDV06 2-8oC Allow samples and sDH2O to adjust to room temperature. Add sDH2O to the vials as follows:

Sample 1: 250 µL

Sample 2: 250 µL

Sample 3: 250 µL

Sample 4: 250 µL

Sample 5: 250 µL

Sample 6: 250 µL

 Test immediately upon receipt/

reconstitution

 ELISA

HA/HI

 R.E.F 39-45 ODs

Titres

 2 rep
Salmonella enteritidis – Abs

JGKPT44

 6 x FD sera (poultry) SE01 – SE06 2-8oC Allow samples and sDH2O to adjust to room temperature. Add sDH2O to the vials as follows:

Sample 1: 200 µL

Sample 2: 200 µL

Sample 3: 200 µL

Sample 4: 200 µL

Sample 5: 200 µL

Sample 6: 200 µL

 Test immediately upon receipt/

reconstitution

 ELISA

RPAT

 R.E.F 39-45 Positive / Negative 1 rep
Mycoplasma gallisepticum - Abs

JGKPT45

 6 x FD sera (poultry) MG01 – MG06 2-8oC Allow samples and sDH2O to adjust to room temperature. Add sDH2O to the vials as follows:

Sample 1: 200 µL

Sample 2: 250 µL

Sample 3: 250 µL

Sample 4: 200 µL

Sample 5: 200 µL

Sample 6: 200 µL

 Test immediately upon receipt/

reconstitution

 ELISA

RPAT

 R.E.F 39-45 Positive / Negative 1 rep
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

None

 

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 22ND of October 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