Method Author VIDAS KUBE Presentation

First page 1 Next page Last page

VIDAS ® KUBE ™ Validation Studies for AOAC OMA & PTM Methods

July 2022 – Patrick Bird & Ronald Johnson

Strictly confidential

P IONEERI NG DI AGNOST I CS

STRICTLY CONFIDENTIAL

NEW GENERATION

• VIDAS KUBE - new VIDAS system to replace VIDAS ® LEGACY systems

• VIDAS ® PC • Mini VIDAS ®

B I O M É R I E U X

STRICTLY CONFIDENTIAL

SYSTEM DESCRIPTION

USER INTERFACE

1 MODULE = 2 SECTIONS x 6 POSITIONS

CLUSTER OF 6 MODULES = 72 POSITIONS

B I O M É R I E U X

STRICTLY CONFIDENTIAL

VIDAS ® KUBE ™ VS VIDAS ® LEGACY

• Compatibility with current VIDAS ® Industry test kits : • Same SPR / strips / calibrators and controls

• Same validated method protocols: • Enrichment, detection and confirmation

• Same technical specifications: • TV calculation, thresholds, instrumental protocols, volumes and pump speed adjustment

• Package inserts

• Optic reader (450 nm)

• SPR blocks at 37 ° C

• Trays at 37 ° C

B I O M É R I E U X

STRICTLY CONFIDENTIAL

OMA AND PTM METHODS IMPACTED

Full Method name

Abbreviation PTM

OMA

VIDAS Listeria monocytogenes 2

LMO2

None

2004.02

VIDAS Listeria monocytogenes Xpress

LMX

091103

2013.11

VIDAS Listeria DUO

LDUO

100702

none

999.06, 2004.06 2010.02

VIDAS Listeria

LIS

981202

VIDAS Listeria Xpress

LSX LPT

None None

VIDAS UP Listeria

2013.1

996.08, 2004.03

VIDAS Salmonella

SLM

None

VIDAS Easy Salmonella

SLM

020901 071101 060903 051201 121806

2011.03 2013.01

VIDAS UP Salmonella

SPT

VIDAS UP E. coli O157:H7

ECPT

none none

VIDAS Campylobacter

CAM

GENE-UP EHEC (VIDAS ESP1 and ESP2)

EHEC

2020.06

B I O M É R I E U X

STRICTLY CONFIDENTIAL

EQUIVALENCE STUDY DESIGN

• Objective : • To ensure that the new VIDAS® KUBE ™ produces equivalent results to the VIDAS ® LEGACY instruments • Systems :

• 3 VIDAS ® KUBE ™ and 3 VIDAS ® LEGACY (2 VIDAS ® PC and 1 mini VIDAS ® )

• 2 Protocols :

• Detection Method • Concentration method for GENE-UP EHEC PCR Assay

• 2 Studies performed for each protocol: • Pure culture • Matrix

B I O M É R I E U X

STRICTLY CONFIDENTIAL

PURE CULTURE STUDY – DETECTION

Assay Name

Strains

Reference

Origin

Listeria monocytogenes 1/2b

ATCC 51780

Dairy products - cheese

• 7 VIDAS Assays Evaluated

Listeria monocytogenes 4b

ATCC 19115

Human

VIDAS LMO2

Listeria ivanovii

ATCC 700402

Unknown

Staphylococcus aureus

ATCC 12600

Pleural fluid

Listeria monocytogenes 1/2b

ATCC 51780

Dairy products - cheese

Listeria monocytogenes 4b

ATCC 19115

Human

• 48 replicates/assay

VIDAS LMX

Listeria ivanovii

ATCC 700402

Unknown

Staphylococcus aureus

ATCC 12600

Pleural fluid

• 2 target strains (10x LOD) • 2 non-target strains

Listeria monocytogenes 1/2b

ATCC 51780

Dairy products - cheese

Listeria innocua

ATCC 33090 ATCC 12600 ATCC 11778

Brain - Cattle Pleural fluid

VIDAS LPT

Staphylococcus aureus

Bacillus cereus

Unknown

Salmonella Typhimurium Salmonella Enteritidis

NCTC 74

Feces - Human

CRA0904090 CRA0906013

Poultry

• 3 VIDAS KUBE

VIDAS SLM

Citrobacter freundii

Vegetables

Enterobacter sakazakii

ATCC 51329

Human

Salmonella Typhimurium

NCTC 74

Feces - Human

• 3 VIDAS Legacy

Salmonella Enteritidis

CRA0904090

Poultry

VIDAS SPT

Citrobacter freundii

CRA0906013

Vegetables

Enterobacter sakazakii

ATCC 51329

Human

• 2 Consecutive Days

Escherichia coli O157:H7

ATCC 43889

Coproculture

Minced meat / ground meat

Escherichia coli O157:H7

ATCC 43895

VIDAS ECPT

• 96 Results/strain

Escherichia coli

ATCC 25922 ATCC 11778

Human

Bacillus cereus

Unknown

Campylobacter jejuni

NCTC 11392

Feces - Human

Campylobacter coli

ATCC 49941

Unknown

VIDAS CAM

Helicobacter pilori

ATCC 43504

Gastric Fluid

Escherichia coli

ATCC 25922

Human

B I O M É R I E U X

STRICTLY CONFIDENTIAL

PURE CULTURE STUDY – DETECTION RESULTS

• Statistical comparison of the test value between 3 VIDAS KUBE and 3 VIDAS Legacy instruments

• Mean, CV and Variance calculated

• Acceptance criteria of <13% CV (internal specification for assays)

• Target Organisms

• 44-48 comparisons <13% CV • 4 Instances outside of acceptance criteria, VIDAS KUBE demonstrated higher repeatability and less variance

• Non-Target Organisms

• All 48 comparisons <13% CV

B I O M É R I E U X

STRICTLY CONFIDENTIAL

PURE CULTURE STUDY – TARGET STRAIN RESULTS

VIDAS ® KUBE™

VIDAS ® LEGACY

VARIANCE

INTERPRETATION

VIDAS ® PARAMETER

POSITIVE STRAIN

TV THRESHOLD

TV MEAN

INTERP. TV CV% VARIANCE

FINAL STATUS

KUBE VS LEGACY

KUBE VS V IDAS 3 0

KUBE VS MINI V IDAS

NCTC 11932 60

0.68 POSITIVE 3.7 0.00060 0.70 POSITIVE 3.5 0.00060

PASSED

CAM

≥ 0,10

ATCC 49941 60

0.99 POSITIVE 2.2 0.00050 1.00 POSITIVE 2.5 0.00060

PASSED

0904090 48

1.20 POSITIVE 3.8 0.00200 1.24 POSITIVE 4.1 0.00260

PASSED

SPT

≥ 0,25

NCTC 74 48

0.94 POSITIVE 2.9 0.00080 0.92 POSITIVE 2.7 0.00060

PASSED

ATCC 43889 48

0.24 POSITIVE 4.1 0.00010 0.25 POSITIVE 4.2 0.00010

PASSED

ECPT

≥ 0,04

ATCC 43895 48

0.25 POSITIVE 4.1 0.00010 0.26 POSITIVE 5.4 0.00020

PASSED

ATCC 51780 48

1.04 POSITIVE 2.9 0.00090 1.00 POSITIVE 2.5 0.00060

PASSED

LPT

≥ 0,05

ATCC 33090 48

1.05 POSITIVE 3.7 0.00150 1.01 POSITIVE 4.6 0.00210

PASSED

0904090 48

0.45 POSITIVE 2.0 0.00008 0.43 POSITIVE 2.7 0.00011

PASSED

SLM

≥ 0,23

NCTC 74 48

0.35 POSITIVE 3.1 0.00011 0.34 POSITIVE 2.3 0.00009

PASSED

ATCC 51780 48

1.12 POSITIVE 3.2 0.00130 1.08 POSITIVE 3.5 0.00142

PASSED

LMO2

≥ 0,05

ATCC 19115 48

0.80 POSITIVE 3.1 0.00063 0.81 POSITIVE 4.8 0.00146

NOK

PASSED

ATCC 51780 48

0.75 POSITIVE 2.1 0.00024 0.73 POSITIVE 2.9 0.00047

NOK

PASSED

LMX

≥ 0,05

ATCC 19115 48

0.78 POSITIVE 1.7 0.00018 0.66 POSITIVE 2.5 0.00027

PASSED

TV- Test value threshold; CV – Correlation of variance, OK – Acceptable data (CV <13%); NOK – Not acceptable data (CV >13%)

B I O M É R I E U X

STRICTLY CONFIDENTIAL

PURE CULTURE STUDY – NON-TARGET STRAIN RESULTS

VIDAS ® KUBE™

VIDAS ® LEGACY

VARIANCE

INTERPRETATION

VIDAS ® PARAMETER

NEGATIVE STRAIN

TV THRESHOLD

TV MEAN

TV CV%

TV MEAN

TV CV%

FINAL STATUS

INTER

VARIANCE

INTER

VARIANCE

KUBEVS LEGACY

KUBEVS VIDAS 30

KUBEVS MINI VIDAS

ATCC 25922 60 ATCC 43504 60 ATCC 51329 48 ATCC 43504 48 ATCC 25922 48 ATCC 11778 48 ATCC 12600 48 ATCC 11778 48 ATCC 43864 48 ATCC 51329 48 ATCC 12600 48 ATCC 700402 48 ATCC 12600 48 ATCC 700402 48

0.02 NEG 0 0.04 NEG 0 0.02 NEG 10 0.02 NEG 20 0.00 NEG 0 0.00 NEG 0 0.00 NEG 0 0.00 NEG 0 0.01 NEG 53 0.01 NEG 14 0.02 NEG 27 0.03 NEG 13 0.00 NEG 0 0.00 NEG 101

0.00000 0.00000 0.00002 0.00204 0.00000 0.00000 0.00000 0.00000

0.02 NEG 32 0.04 NEG 0 0.02 NEG 0 0.05 NEG 18 0.00 NEG 0 0.00 NEG 0 0.00 NEG 0 0.00 NEG 0 0.01 NEG 0 0.01 NEG 0 0.02 NEG 210 0.02 NEG 0 0.01 NEG 0 0.01 NEG 74

0.00003 0.00000 0.00002 0.00258 0.00000 0.00000 0.00000 0.00000

OK OK OK OK OK OK OK OK OK OK OK OK OK OK

PASSED PASSED PASSED PASSED PASSED PASSED PASSED PASSED PASSED PASSED PASSED PASSED PASSED PASSED

CAM

< 0,10

SPT

< 0,25

ECPT

< 0,04

LPT

< 0,05

Siegel-Tukey, Ansari Bradley, Klotz and

SLM

< 0,23

0.00003 0.00001 0.00000 0.00003

0.00004 0.00004 0.00000 0.00002

LMO2

< 0,05

LMX

< 0,05

TV- Test value threshold; CV – Correlation of variance, OK – Acceptable data (CV <13%); NOK – Not acceptable data (CV >13%)

B I O M É R I E U X

STRICTLY CONFIDENTIAL

VIDAS ESPT (ESP1 AND ESP2) CONCENTRATION

• GENE-UP ® EHEC method :

Day2

Day1

8-18h*

10-20h

12-22h

13-23h

31-45h

VIDAS® ESPT : O26, O45, O103 O111, O121, O145, O157

PCR Stx1&2/eae

PCR O26, O45, O103, O111, O121, O145

PCR O157:H7

ChromID® EHEC ChromID ® E. coli

Slidex®

VIDAS® ESP1

VIDAS® ESP2

B I O M É R I E U X

STRICTLY CONFIDENTIAL

PURE CULTURE STUDY – CONCENTRATION

5 TARGET STRAINS 6 REPETITIONS - ESP1 PROTOCOL 6 REPETITIONS - ESP2 PROTOCOL 3 VIDAS ® LEGACY SYSTEMS + 3 VIDAS ® KUBE ™ SYSTEMS = 360 VIDAS ® ESPT PREPARATIONS

VIDAS ® LEGACY

VIDAS ® KUBE ™

CP comparison

EHEC Numeration

CP comparison

EHEC Numeration

B I O M É R I E U X

STRICTLY CONFIDENTIAL

PURE CULTURE RESULTS - CONCENTRATION

Test Brown Forsythe

Normality test

Variance analysis

Kruskal Wallis

Analysis Variable : CP_

Acceptance Criteria CP Mean max +/- 2 CP

Statistical equivalence

FINAL STATUS RESULT

Strain

PCR

Std Dev

Coeff of Variation

Instrument N Obs Mean

Median Min Max Range

Pvalue

VIDAS KUBE

O111

18 33.54 0.25 33.56 32.89 33.94 1.05

0.76

EH1

>0.05

0.2831 <0.0001

NOK

CONFORM CONFORM

VIDAS LEGACY

CRA 0905094

18 32.90 0.17 32.96 32.59 33.22 0.63

0.53

VIDAS KUBE

O26

18 33.71 0.42 33.81 33.08 34.67 1.59

1.25

EH1

>0.05

0.2099

0.5238

CONFORM CONFORM CONFORM

VIDAS LEGACY

CRA 0905088

18 33.64 0.27 33.63 33.06 34.14 1.08

0.81

VIDAS KUBE

O145

18 34.60 0.17 34.56 34.21 34.90 0.69

0.49

EH1

>0.05

0.0268

0.0006

NOK

CONFORM CONFORM

VIDAS LEGACY

CRA 0905089

18 34.20 0.31 34.18 33.54 34.72 1.18

0.92

VIDAS KUBE

O157

18 32.12 0.44 32.15 31.22 32.96 1.74

1.38

EH1

>0.05

0.0330

0.9748

CONFORM CONFORM CONFORM

VIDAS LEGACY

CRA 9405023

18 32.11 0.26 32.12 31.64 32.66 1.02

0.81

VIDAS KUBE

O157

18 31.28 0.26 31.28 30.89 31.87 0.98

0.83

ECO

>0.05

0.3480

0.5978

CONFORM CONFORM CONFORM

VIDAS LEGACY

CRA 0905023

18 31.32 0.21 31.33 30.93 31.68 0.75

0.66

VIDAS KUBE

O103

18 28.55 0.39 28.37 28.06 29.34 1.28

1.36

EH2/ TOP6

<0.05

0.2680

CONFORM CONFORM CONFORM

VIDAS LEGACY

CRA 0905012

18 28.28 0.46 28.47 26.93 28.79 1.86

1.61

VIDAS KUBE

O111

18 28.30 0.47 28.24 27.61 29.69 2.08

1.68

EH2/ TOP6

<0.05

0.4571

CONFORM CONFORM CONFORM

VIDAS LEGACY

CRA 0905094

18 28.36 0.40 28.30 27.67 28.97 1.30

1.40

B I O M É R I E U X

STRICTLY CONFIDENTIAL

PURE CULTURE RESULTS - CONCENTRATION

Test Brown Forsythe

Normality test

Variance analysis

Kruskal Wallis

Analysis Variable : CP_

Acceptance Criteria CP Mean Ecart max +/- 2 CP

FINAL STATUS RESULT

Statistical equivalence

Strain

PCR

Coeff of Variation

Instrument

N Obs Mean Std Dev Median Min Max Range

Pvalue Pvalue Pvalue Pvalue

VIDAS KUBE

O111

18 44.56 39.15 36.50 1.00 111.00 110.00 87.86

EH1

>0.05 0.0025 0.4569

CONFORM CONFORM CONFORM

CRA 0905094

VIDAS LEGACY

18 28.22 16.66 32.00 2.00 60.00 58.00

59.02

VIDAS KUBE

O26

18 100.94 59.35 126.00 2.00 193.00 191.00 58.80

EH1

>0.05 0.5685 0.1670

CONFORM CONFORM CONFORM

CRA 0905088

VIDAS LEGACY

18 74.61 52.30 66.50 6.00 182.00 176.00 70.10

VIDAS KUBE

O145

18 118.33 101.20 79.50 11.00 300.00 289.00 85.52

EH1

<0.05

0.2415 CONFORM CONFORM CONFORM

CRA 0905089

VIDAS LEGACY

18 145.94 69.16 143.50 7.00 300.00 293.00 47.39

VIDAS KUBE

O157

18 140.00 27.97 138.50 96.00 192.00 96.00

19.98

EH1

>0.05 0.7855 0.0538

CONFORM CONFORM CONFORM

CRA 9405023

VIDAS LEGACY

18 158.83 28.60 159.50 104.00 206.00 102.00 18.01

VIDAS KUBE

O103

18 215.67 88.32 235.50 34.00 300.00 266.00 40.95

EH2/ TOP6

<0.05

0.1379 CONFORM CONFORM CONFORM

CRA 0905012

VIDAS LEGACY

18 255.83 56.32 285.50 134.00 300.00 166.00 22.01

B I O M É R I E U X

STRICTLY CONFIDENTIAL

MATRIX STUDY – DETECTION

• 4 VIDAS assays

• Minimum 30 food items

• 2 Target strains

• 1 Non-target strain

• 3 VIDAS KUBE and 3 VIDAS LEGACY Instruments

• Comparison of Test Values between instruments

• VIDAS KUBE mean should be within ± 20% VIDAS LEGACY mean TV

• Calibrators and controls conform to MLE acceptance criteria

B I O M É R I E U X

STRICTLY CONFIDENTIAL

MATRIX STUDY RESULTS – CALIBRATORS & CONTROLS

• Acceptance criteria met for MLE • S1 TV mean within [Min S1 RFV; Max S1 RFV] • S1 RFV CV% < 13% • C1 TV within [Min C1 TV; Max C1 TV] • C2 TV < MLE C2 TV

B I O M É R I E U X

STRICTLY CONFIDENTIAL

MATRIX STUDY RESULTS – FOOD STUDY

• Acceptance criteria met for food study • TV Mean within Acceptance Criteria

B I O M É R I E U X

STRICTLY CONFIDENTIAL

MATRIX STUDY RESULTS – FOOD STUDY

• No Negative or Positive Deviations

B I O M É R I E U X

STRICTLY CONFIDENTIAL

PURE CULTURE STUDY - CONCENTRATION

20 negative + spiked food samples 2 instrumental protocols ESP1 and ESP2 3 VIDAS ® LEGACY SYSTEMS + 3 VIDAS ® KUBE ™ SYSTEMS = 480 VIDAS ® ESPT preparations

VIDAS ® LEGACY

VIDAS ® KUBE ™

CP comparison

EHEC Confirmation

B I O M É R I E U X

STRICTLY CONFIDENTIAL

MATRIX STUDY – CONCENTRATION

Food sample

Protocol

Food sample number

25g or 375g of sample

225 mL BWP for weight 1 or 1125mL for weight 2 Mix with a paddle blender incubate at 41°C ± 1 °C for 8-24 hours for weight 1 or 10-24 hours for weight2

Raw meat (except poultry) – weight 1: 25g weight 2: 375g

X g (or X mL) of sample

9X mL of BWP Mix with a paddle blender incubate at 37°C ± 1 °C for 18-26 hours

Raw milk products (up to 25g or 25mL)

X g of samples

9X mL of BWP Mix with a paddle blender incubate at 41°C ± 1 °C for 18-24 hours

Vegetables (up to 25g)

TOTAL

B I O M É R I E U X

STRICTLY CONFIDENTIAL

MATRIX STUDY – CONCENTRATION

Mean bias and confidence interval at 95% from bias

Acceptance Criteria CP Mean Ecart max +/- 2 CP

Number of food samples

VIDAS® KUBE ™

VIDAS® LEGACY

PCR

Signification

FINAL STATUS

Bias ≠ 0

EH1 PCR eae detection

0,09 [-0,14 ; 0,32]

significative difference between both system

34,34

34,25

CONFORM

Bias ≠ 0

EH1 PCR stx1/stx2 detection

0,07 [-0,20 ; 0,34]

significative difference between both system

30,59

30,51

CONFORM

Bias ≠ 0

0,17 [-0,41 ; 0,76]

significative difference between both system

ECO PCR

31,97

31,8

CONFORM

Bias and interval > 0 => VIDAS ® KUBE ™ > VIDAS ® LEGACY

0,85 [0,27 ; 1,43]

TOP6 PCR

32,6

31,75

CONFORM

B I O M É R I E U X

STRICTLY CONFIDENTIAL

MATRIX STUDY – CONCENTRATION

VIDAS® KUBE™ 1

VIDAS® KUBE™ 2

VIDAS® KUBE™ 3

miniVida s

VIDAS PC 1

VIDAS PC 2

date

Food sample

Strain

Concentration

Acceptance criteria COMPARISON

17/06/2022 17/06/2022 17/06/2022 22/06/2022

beef raw meat

5.10^5 UFC/ml 5.10^5 UFC/ml 5.10^5 UFC/ml 5.10^5 UFC/ml 5.10^5 UFC/ml 5.10^5 UFC/ml 5.10^5 UFC/ml 5.10^5 UFC/ml 5.10^5 UFC/ml 1.10^5 UFC/ml 1.10^5 UFC/ml 1.10^5 UFC/ml 1.10^5 UFC/ml

CH CH CH CH CH CH BH BH BH BH CH BH CH BH CH CH CH BH CH BH CH BH CH CH CH CH CH DH DH CH CH CH CH DH DH CM CH CM CMCH DH DH CM CM CM CM CM CM BH BH BH BH BH BH BMBH BMBH BMBH BMBH BMBH bh DH DH DH DH DH DH CM BM CM BM BM BM CM CM CM CM CM CM CH BMBH BH CH CH CH

CONFORM CONFORM CONFORM CONFORM CONFORM CONFORM CONFORM CONFORM CONFORM CONFORM CONFORM CONFORM

O157: CRA 9405023

goat raw milk cheese

cucumber

beef raw meat

O26 : CRA 0905088

22/06/2022 roquefort lamb raw milk cheese

22/06/2022 22/06/2022

rawspinash

Pork raw meat

O145: CRA 0905089

22/06/2022 brie cow raw milk cheese

22/06/2022 24/06/2022 24/06/2022 24/06/2022 24/06/2022

iceberg lettuce cow raw milk Pork raw meat Lamb's lettuce beef carpaccio

O111: 0905094 O111: 0905094 O111: 0905094 O103 : 0905012

Equivalent confirmation

24/06/2022 munster cow raw milk cheese

O103 : 0905012

1.10^5 UFC/ml

BH BH BH BH BH BH

CONFORM

24/06/2022 29/06/2022 29/06/2022

frozenspninach Lamb raw meat Veal raw meat

O103 : 0905012 O26: 1301130 O103: 1301170

1.10^5 UFC/ml 1.10^6 UFC/mL 5.10^5 UFC/ml

AM AM AM AM AM

CONFORM CONFORM CONFORM

BM BM BM BM BM BM CH CH CH CH CH BH

camembert cow raw milk cheese

29/06/2022

O111 : 1103167 --> O26 1.10^6 UFC/ml

CH CH CH CH CH CH

CONFORM

29/06/2022 cantal cow raw milk cheese

O145 : 1301171

5.10^6 UFC/ml

CH CH CH CH CH CH

CONFORM

29/06/2022 St Félicien cow raw milk cheese

O157 : 9405025 5.10^5 UFC/mL

DH DH DH DH DH DH

CONFORM

B I O M É R I E U X

STRICTLY CONFIDENTIAL

CONCLUSION

• Equivalent detection or concentration after VIDAS ® KUBE ™ preparation

• Acceptance criteria were met for each study • Calibrators and controls were within MLE acceptance criteria

• TV Mean of VIDAS KUBE within acceptance interval • Evaluation of food matrices produced equivalent results • No positive or negative deviations • Confirmation on specific agar is equivalent between prepared samples with VIDAS ® KUBE ™ and VIDAS ® LEGACY.

B I O M É R I E U X

P I O N E E R I N G D I A G N O S T I C S

Page 2 Page 3 Page 4 Page 5 Page 6 Page 7 Page 8 Page 9 Page 10 Page 11 Page 12 Page 13 Page 14 Page 15 Page 16 Page 17 Page 18 Page 19 Page 20 Page 21 Page 22 Page 23 Page 24

Made with FlippingBook Annual report maker