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AdGène laboratoire 1 rue des conquérants – 14220 Thury-Harcourt – www.adgene.fr
Email : [email protected] / tel : 02 31 15 62 80 / fax : 02 31 15 62 85
BIO-RAD 3 boulevard Raymond Poincaré 92430 MARNES LA COQUETTE
NF Validation
Validation of alternative‘s analysis method for Legionella spp
detection and quantification
Renewal study for iQ-Check® Legionella spp. for the detection and enumeration of
Legionella in sanitary hot water and cooling towers
N° agreement BRD 07/15 – 12/07
Summary Report
This study include 15 pages whose 0 annexe. Reproduction of this report is only authorized in its integral form.
Version 1.1
14 september 2016 Cancels and replaces the previous version The old version must be destroyed.
2 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Summary
Reminder of the method of analysis ....................................................................................................... 3
Date of first attribution of NF Validation trade mark .......................................................................... 3
Method principle ................................................................................................................................. 3
Protocol references: ............................................................................................................................ 4
Application domain ............................................................................................................................. 4
Restrictions .......................................................................................................................................... 4
Methods of reference ......................................................................................................................... 4
Validation history .................................................................................................................................... 5
Summary of results.................................................................................................................................. 7
Linking of calibration solution and reference material to primary standard ...................................... 7
Calibration function ............................................................................................................................. 8
Validation of limit of detection ........................................................................................................... 8
Validation of limit of quantification .................................................................................................. 10
Study of efficiency and robustness of extraction .............................................................................. 11
Selectivity: inclusivity and exclusivity ................................................................................................ 12
Inclusivity ....................................................................................................................................... 12
Exclusivity ...................................................................................................................................... 12
Practicability ...................................................................................................................................... 12
Ease of use: .................................................................................................................................... 12
Fast results report: ........................................................................................................................ 12
Results security: ............................................................................................................................. 12
Inter-laboratories study .................................................................................................................... 13
Bibliography study ................................................................................................................................. 14
Overview of customer reclamation ....................................................................................................... 14
Conclusions ............................................................................................................................................ 15
3 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Reminder of the method of analysis
Date of first attribution of NF Validation trade mark iQ Check® Legionella spp. and iQ Check® Legionella pneumophila kits was validated to allow
utilization of trademark NF VALIDATION in 2007. Authorization was renewed in 2011, follows
by two extensions in 2012 (see chapter Validation History).
Method principle iQ Check® Legionella spp kit is intended to detect or to quantify bacteria genius Legionella in
water sample, due to Polymerase Chain Reaction (PCR). PCR allows amplification and
detection of specific sequences with specific primers and fluorescent probe.
Principle is based on three steps (see Annex 1):
- Sample filtration
- DNA extraction with Aquadien kit (and protocol W2 for clogging samples).
- Legionella spp target sequences amplification.
DNA extraction with Aquadien kit is based on alkaline lysis wih thermal choc. It is followed by
an ultrafiltration purification step. A DNA fraction is amplified by real-time PCR.
Primers hybridize to target sequence during PCR reaction. Taq polymerase use primers and
triphosphates desoxynucleotides (dNTPs) to stretch DNA and to create copies of Legionella
spp target DNA.
Specific probe hybridize to amplicons during PCR. This probe is labelled with a fluorophore
which emit fluorescence only after hybridization. Fluorescence intensity increases
proportionally with increasing of PCR products.
Fluorescence is directly measured by optical machinery of the thermocycler during
hybridization step. Thermocycler software cast in real-time the measured fluorescence function
of number of amplification cycles. Software determines a Ct (cycle from which fluorescence is
higher than background signal). Reading Ct permits to detect presence of Legionella spp target
sequences. Detection of target sequences indicates presence of the bacteria in analysed water
sample.
Quantification is possible by using calibrated DNA solutions calibré iQ Check® Legionella
Quantification Standards. These standards are connected to primary standard of Centre
National de Référence des Légionelles.
PCR inhibition phenomenon is detected by utilisation of a synthetic DNA (internal control –
IPC) included in amplification solution with each sample. IPC is amplified during same time
than target sequences, with same primers but with a different probe and a different fluorphore.
4 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
iQ Check® Legionella spp. kits are validated with following materials:
Software Opticon Monitor 3.4
CFX manager Software Industrial Diagnostic Edition v2.2
CFX manager Software Industrial Diagnostic Edition V2.2
Chassis PTC-200 C1000 C1000 touch Optical Head / heating block Chromo4 CFX96 CFX96 Deep
well touch
Protocol references: Aquadien™ (Réf. 3578121) : 10/2013 – Code : 881116
iQ-Check® Legionella spp (Réf. 3578102) : 10/2013 – Code : 881117
Application domain Samples of all types of water.
Restrictions Certification of the kit deals with it utilization with thermocyclers Bio-Rad Chromo™4 and
CFX96 Deep well Touch.
Methods of reference NF T90-471 (juin 2015) : Qualité de l’eau - Détection et quantification des Legionella et/ou
Legionella pneumophila par concentration et amplification génique par réaction de
polymérisation en chaîne en temps réel (RT-PCR).
ISO/TS 12869 (November 2012) : Water quality -- Detection and quantification of Legionella
spp. and/or Legionella pneumophila by concentration and genic amplification by quantitative
polymerase chain reaction (qPCR)
PCR Validation protocol:
Protocole de validation pour les méthodes commerciales de détection et de dénombrement
de Legionella et Legionella pneumophila par concentration et amplification génique par
réaction de polymérisation en chaîne (PCR) V3.0
5 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Validation history
Certified method Certificate Date
approval Object Commentaries
Expert laboratory
PCR validation protocol
iQ-check Legionella spp.
BRD 07/15-12/07
18/12/2007 Validation initial IPL SED
Nord Rev. 0 (2006)
10/06/2011 Renewal 1
Mix PCR evolution 2 extractions method
(protocol W2) Update according to protocol version 1
IPL SED Nord
Rev. 1 (2011)
04/04/2012 Extension 1 New thermocycler (Deep Well touch)
Eurofins IPL Nord
Rev. 1 (2011)
27/05/2013 Extension 2 Validation protocol
V.2 NA
Rev. 2 (2013)
05/11/2013 Modification Software V2.1 NA Rev. 2 (2013)
09/03/2015 Modification Software V2.2 NA Rev. 2 (2013)
18/12/2015 Renewal 2
AdGène with
extension on
qualitative test
Rev 3 (2015)
2007:
Method was validated initially in 2007.
2011:
2010/211 study for renewal of validation considered the modifications of validated kit and
of validation protocol (renewal n°1considering norm NT T90-471 published in April 2010).
A third party study has focused on two first phases of validation protocol aiming to verify
supplier announced performances for new formulation of iQ-Check® L. spp kit:
Phase 1: Study of limit of detection and limit of quantification of PCR step, calibrating
function, link to primary standard, efficiency and robustnessof extraction with Aquadien™
kit. New thermocycler CFX 96 was implemented.
Phase 2: Study of inclusivity and of exclusivity, of practicability and of reagents quality.
Interlaboratory study realised in 2007 was not made again
New modification from initial validation was :
- iQ-Check® L. spp kit: New origin of Taq polymerase and chemical evolution of IPC
probe (TEXAS RED fluorophore was replaced by HEX fluorophore)
- Aquadien Kit: two modalities of utilization according to sample filterability (protocol
W2 for clogging samples added to classical protocol) and horizontal double- tangential microfiltration for DNA purification step. Membranes and materials
composition do not change.
New thermocycler can be used: CFX96 with CFX Manager Software Industrial
Diagnostic Edition version V1.1.
6 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
2012:
Validation extension was pronounced in 2012 after evolution of characteristics of
thermocycler CFX96 which becomes CFX96 Deep Well Touch. Modifications concern
reactional volume of heating block, user interface (keyboard and screen), and software
CFX Manager which pass in version V1.2
AFNOR Certification Technical office qualified theses evolutions as minority and without
impact on kit performance. No new assays were performed.
2013:
Late May 2013: Validation of iQ-Check® L. spp method was extended to norm ISO/TS
12869. No study complement was necessary: Assays performed according to norm NF
T90-471 answers to requirements of ISO/TS 12869 and follow migration to revision 2 of
validation protocol.
November 2013: Evolution of software CFX manager IDE v2.1. No study complement
was necessary.
2015:
March 2015: Evolution of software CFX manager IDE v2.2. No study complement was
necessary.
October 2015: Renewal of iQ-Check® L. spp method with extension on detection
(qualitative research) of Legionella spp without supplementary test. AFNOR Certification
Technical office qualified this evolution without impact on kit performance. No new
assays were performed.
7 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Summary of results Next results were obtained with revision V1.0 of validation protocol for commercial method of
detection and quantification of Legionella and Legionella pneumophila by concentration and
gene amplification by Polymerase Chain Reaction (PCR). Results are conforms with version
V2.0 because it contains no major modification causing complementary study.
Linking of calibration solution and reference material to primary standard
Methodology
Linking of working calibration solution to primary standard is made to cover the quantification
domain with 3 ranges of calibrated DNA iQ-Check® Legionella spp. which contain 4 levels of
concentrations of Genome Unity of Legionella pneumophila serogroup (QS1, QS2, QS3, QS4)
and 3 independent ranges of primary standard aiming at the 4 levels of concentrations of range
of calibrated DNA iQ-Check® Legionella Quantification Standards.
Linking of reference material to primary standard is evaluated analysing results of 2 deposits
of reference material given with iQ-Check® Legionella spp. kit.
Results
Analysed parameters for evaluation of linking of calibration solution and of reference material
to primary standard on thermocycler CFX96 and chromo 4 are submitted in next table:
Regression curve Correlation Efficiency (%)
Reference range (CFX96) C(t) average = -3.198.log(x) + 39.076 0.998 105.5
Reference range (Chromo 4) C(t) average = -2.891.log(x) + 38.674 0.995 121.75
Calibration solution Calibration error
QS1 QS2 QS3 QS4
Per level (CFX96) 0.07 0.20 0.14 0.07
Per level (Chromo 4) 0.03 0.30 0.23 0.16
Average (CFX96) 0.12
Average (Chromo 4) 0.18*
Slopes equivalence (CFX96) 0.00
Slopes equivalence (Chromo 4) 0.13
Reference material Calibration error
CFX96 0.19
Chromo 4 0.19
* Calibration error of calibration solution is 0.18log with thermocycler Chromo4. However, equivalence of slopes from reference range and calibration solution range is verified.
Calibration error of calibration solution is lower than 0.15log. Slopes from reference range and calibration solution range are equivalent. Conclusion
8 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Calibration solution and reference material of iQ-Check® Legionella spp kit satisfy conditions
of linking to primary standard with thermocycler CFX96.
Calibration solution globally satisfies conditions of linking to primary standard with
thermocycler Chromo 4. Reference material of iQ-Check® Legionella spp kit satisfies
conditions of linking to primary standard with thermocycler Chromo 4.
Calibration function
Methodology
Study of calibration function is made deposit 5 different reference ranges of calibrated DNA
solution iQ-Check® Legionella Quantification Standards (comprising 4 levels of concentration
of Genome Unity of Legionella pneumophila), given with iQ-Check® Legionella spp. kit.
5 measures are made with iQ-Check® Legionella spp kit for each level of concentration in
reproducibility conditions.
Results
Equation of regression curve and efficiency of PCR reaction are defined in these conditions.
Results are obtained on CFX96.
Regression curve -3.197.log(x) + 41.347
Efficiency 105.5%
Performances of linear regression:
QS1 QS2 QS3 QS4
Bias 0.06 -0.10 0.00 0.04
Standard deviation 0.12 0.06 0.08 0.05
Exactitude of linearity 0.13 0.12 0.08 0.07
uncertainty of linearity 0.42 0.37 0.27 0.22
Conclusion
Linear regression satisfies exigence of exactitude lower than 0.15log for each level of reference
range. Linearity is verified on the whole domain cover by the range of calibrated DNA solution
iQ-Check® Legionella Quantification Standards given with iQ-Check® Legionella spp. kit.
Validation of limit of detection
Methodology
9 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Evaluation of limit of detection is made from 30 independent dilutions of Legionella
pneumophila DNA in concentration of 5UG per PCR reaction. Duplicate amplifications are
made in repeatability conditions. Results are obtained on CFX96.
Results
Conclusion
The 30 duplicate are positives. Limit of detection is validated for 5 UG per PCR reaction.
All Ct in previous table are lower than intercept. Qualitative detection is conforming.
10 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Validation of limit of quantification
Methodology
Evaluation of limit of quantification is made from 30 independent dilutions of Legionella
pneumophila DNA in concentration of 15UG per PCR reaction. Duplicate amplifications are
made in repeatability conditions. Results are obtained on CFX96.
Results
Results Theoretical values or validation criteria
Average x’ (Log UG/reaction) 1.309 1.279 Standard deviation (Log UG/ reaction)
0.097
Bias 0.030 LQ Exactitude 0.101 0.15 LQ Uncertainty 0.207
Conclusion
Value of exactitude of limit of quantification is estimated at 0.101. This value is lower than 0.15.
Limit of quantification is validated for 15 UG per PCR reaction for iQ-Check®Legionella spp.
kit.
Positivity threshold
User manual foresees a Ct of 43 hereafter whose samples are considered as lower than the
limit of detection
All values for characterisation of limit of detection have Ct lower than 43. This value
corresponds to the positivity threshold lower than limit of detection.
11 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Study of efficiency and robustness of extraction
Methodology
Studies of extraction efficiency were realized with extraction kit Aquadien™ (for clean waters)
and Aquadien W2 (for clogging waters). Efficiency was evaluated on 10 independent samples,
which were artificially contaminated with two levels of concentrations of Legionella
pneumophila ATCC 33152 (1000 and 100 000 UG / PCR reaction). Samples were 3 different
matrixes: sterile water, domestic hot water and water from air cooling-tower.
Samples were artificially contaminated by primary bacterial suspension. It concentration was
determined by 3 quantifications after an extraction step of DNA by direct lysis on 3 aliquots.
Results are obtained on CFX96.
Results
Extraction efficiency
Aquadien Protocol Aquadien W2 Protocol
Log Average Log Average
Domestic hot water 1000 UG/L 100 000 UG/L
-0.29 -0.39
-0.34 -0.16 -0.45
- 0.30
Water from air cooling-tower
1000 UG/L 100 000 UG/L
-0.09 -0.31
-0.20 -0.45 -0.47
- 0.46
Mineral water 1000 UG/L 100 000 UG/L
-0.25 -0.42
-0.33 -0.55 -0.46
- 0.50
Average efficiency (log) -0.29 -0.41 Variance (log) 0.04 0.03 Global extended uncertainty (log) 0.71 0.89
Conclusion
Study of efficiency and robustness of extraction method allows evaluating average efficiency
of:
- Aquadien method: -0.29log
- Aquadien W2 method: -0.41log
Efficiencies with two extraction methods are conforming to criteria -0.6 log / +0.3 log
(equivalent to efficiency comprise between 25% and 199%).
12 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Selectivity: inclusivity and exclusivity DNA was extracted from pure bacterial suspension for each strain.
Inclusivity
Inclusivity assays were realized on DNA extracts with concentration about 100 UG/ PCR
reaction. Concentrations were estimated by O.D.600nm of bacterial suspension.
DNA of 35 strains of tested Legionella (15 Legionella pneumophila et 20 Legionella spp) were
amplified.
Exclusivity
Exclusivity assays were realized on DNA extracts with concentration about 10 000 UG/ PCR
reaction. Concentrations were estimated by O.D.600nm of bacterial suspension.
DNA of 16 strains of tested were not amplified, except 5 of them which show weak
amplification.
Conclusion
iQ-Check® Legionella spp kit selectivity is satisfying.
Practicability
Ease of use: reagents are all supplied with kits and are ready-to-use. Serial analyses from 1
to 30 samples, for quantification, are easy to make. A technician, who knows microbiology and
molecular biology techniques and the specific thermocycler and it software, can be trained in
1 day.
Fast results report: duration of different phases is compatible with a short results report
(5hours).
Results security: It guarantees by utilization of inhibition internal control (in same reaction well
than sample) and by a software of results analysis. Use of software ensures traceability of
complete information.
13 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Inter-laboratories study
Methodology
Inter-laboratories study was realized in 2007 with 14 collaborating laboratories. Results of one
laboratory were not taken into account because of technical problem which invalidated
standardization. 13 laboratories were retained for statistical exploitation.
Goal of this study is to evaluate fidelity (repeatability and reproducibility) of iQ-Check®
Legionella spp. method:
- For only amplification step (2 DNA solutions of L. anisa et L. pneumophila sg1 at 2
different levels of concentration).
- For complete analysis (concentration, lysis, extraction, purification and gene
amplification) on characterized bacterial suspensions of L. pneumophila and
Escherichia coli (CIP 54.8) at 2 different levels of concentration).
- For whole analysis in real situation (hot domestic water naturally contaminated by
L. pneumophila and Legionella spp).
- For a water guarantees without any DNA of Legionella.
Results
Sample types Calibrated DNA
solutions
Contaminated hot
domestic water Natural water
Contamination
levels
(UG/L)
L. pneumophila
ATCC 33152
2000
UG/µl
20000
UG/µl
4000
UG/200
ml
40000
UG/200
ml
Hot domestic
water naturally
contaminated
L. anisa 500
UG/µl
5000
UG/µl
1000
UG/200
ml
10000
UG/200
ml
E. coli
5000
UG/200
ml
50000
UG/200
ml
Number of
laboratories
participating 14 14 14 14 14
retain 13 13 13 13 13
Homogeneity
assay
Analysis number 20 20 9 9 9
Average (Log) 2.91 3.97 3.42 4.41 3.76
Results
Average (Log) 3.02 4.11 3.52 4.47 3.69
r (Log) 0.18 0.15 0.28 0.34 0.46
R (Log) 0.43 0.32 0.72 0.66 0.8
Sr (Log) 0.06 0.06 0.10 0.12 0.16
SR (Log) 0.14 0.10 0.24 0.20 0.23
14 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Conclusion
Repeatability values in r (log) are about 0.15 for DNA solutions (only PCR step) and about 0.7
for bacterial suspensions (global method). This is acceptable. Signification of these results is
that we can wait for factor 2 measurement of deviation in a same laboratory. Repeatability is
not a major source of error.
Reproducibility values in R (log) are about 0.4 for DNA solutions (only PCR step) and about
0.7 for bacterial suspensions (global method). Compared to repeatability, this order of
magnitude is equivalent to values that we can obtain for environmental microbiology analyses.
Signification of these results is that we can wait for factor 5 of measurement deviation between
2 different laboratories. Reproducibility does not participate in a unreasonable way to result
dispersion.
Bibliography study
- No new publication found since the last method publication
- No external validation realized by other certification organism.
Overview of customer reclamation
No customer reclamation.
15 AdGène laboratoire iQ-Check® Legionella spp. summary report V1.1
Conclusions
Performances of iQ-Check® Legionella spp. method are conforming to requirement of norms NF T90-471 and ISO/TS 12869, and of AFNOR validation protocol: “Protocole de validation pour les méthodes commerciales de détection et de dénombrement de Legionella et Legionella pneumophila par concentration et amplification génique par réaction de polymérisation en chaîne (PCR) V3.0”. Modifications between versions 2.0 and 3.0 of AFNOR validation protocol concern positivity threshold (qualitative detection) and do not need complementary study. iQ-Check® Legionella spp kit is a kit validated for detection and for Detection and Quantification of Legionella and/or Legionella pneumophila by concentration and gene amplification by real-time Polymerase Chain Reaction (qPCR).
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
ANNEXE C
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1,75
2
0,18
0,13
38,6
74
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
ANNEXE D
FONCTION DE CALIBRAGE
Fonction de calibrage
Niveau (UG/puits) x i 19 390 3900 39000 19 390 3900 39000x' i =Log(x i ) 1,28 2,59 3,59 4,59 1,28 2,59 3,59 4,59
gamme gamme 1 37,08 33,33 29,60 26,49yij 36,49 33,27 29,84 26,72
gamme 2 37,02 33,18 29,44 26,36k=5 répétitions 36,77 33,33 29,71 26,24
gamme 3 37,78 33,30 29,69 26,5436,69 33,31 29,99 26,46
gamme 4 37,32 33,81 30,14 26,6537,16 33,32 30,32 26,84
gamme 5 37,40 33,57 30,01 26,5236,90 33,41 30,05 26,50
Moyenne mi 37,06 33,38 29,88 26,53 37,06 33,38 29,88 26,53
Estimation de la droite de régression
Pente a = -3,197Ordonnée à l'origine b = 41,347
Estimation de l'efficacité
Efficacité e = 105,5%
Vérification des performances de la régression linéaire
Niveau x i 19 390 3900 39000 19 390 3900 39000x' i =Log(x i ) 1,28 2,59 3,59 4,59 1,28 2,59 3,59 4,59
gamme gamme 1 1,33 2,51 3,67 4,65yij 1,52 2,53 3,60 4,57
gamme 2 1,35 2,55 3,72 4,69k=5 répétitions 1,43 2,51 3,64 4,73
gamme 3 1,12 2,52 3,65 4,631,46 2,51 3,55 4,66
gamme 4 1,26 2,36 3,51 4,601,31 2,51 3,45 4,54
gamme 5 1,23 2,43 3,55 4,641,39 2,48 3,53 4,64
Moyenne mi 1,34 2,49 3,59 4,63 1,34 2,49 3,59 4,63
Biais 0,06 -0,10 0,00 0,04
Ecart type S = 0,12 0,06 0,08 0,05
Exactitude de linéarité ELIN = 0,13 0,12 0,08 0,07
Incertitude de linéarité ULIN = 0,42 0,37 0,27 0,22
1,31 2,46 3,54 4,64
1,28 2,43 3,48 4,57
1,29 2,52 3,60 4,64
1,39 2,53 3,68 4,71
1,43 2,52 3,64 4,61
37,15 33,49 30,03 26,51
37,24 33,57 30,23 26,75
37,24 33,31 29,84 26,50
36,90 33,26 29,58 26,30
36,79 33,30 29,72 26,61
Droite de calibrage
20
22
24
26
28
30
32
34
36
38
40
0,00 1,00 2,00 3,00 4,00 5,00x'i
C(t)
gamme 1
gamme 2
gamme 3
gamme 4
gamme 5
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
ANNEXE E
LIMITE DE DETECTION
Limite de détection à 5UG
Echantillons à la concentration 5UG
Well Sample C(t) I.C. C(t) SQe1 37,88 33,12 6,445
A02 e1 37,73 33,12 7,232B02 e2 39,38 36,36 1,982C02 e2 38,41 34,8 4,257D02 e3 37,65 34,56 7,690E02 e3 38,19 34,14 5,046F02 e4 37,92 34,09 6,237G02 e4 38,12 34,01 5,328H02 e5 38,25 32,97 4,826A03 e5 37,77 33,71 7,028B03 e6 38,99 35,71 2,685C03 e6 38,34 34,12 4,496D03 e7 38,1 34,39 5,432E03 e7 38,02 34,24 5,775F03 e8 39,21 34,53 2,268G03 e8 37,86 34,21 6,526H03 e9 37,39 32,78 9,424A04 e9 37,82 33,79 6,734B04 e10 41,57 39,12 0,356C04 e10 37,97 34,42 5,989D04 e11 38,9 34,49 2,887E04 e11 38,13 34,04 5,271F04 e12 38,93 34,35 2,816G04 e12 37,85 34,08 6,557H04 e13 37,15 32,87 11,400A05 e13 38,08 34,53 5,489B05 e14 38,19 34,68 5,041C05 e14 38,56 34,31 3,786D05 e15 37,91 34,44 6,301E05 e15 38,43 34,93 4,193F05 e16 38,04 34,35 5,667G05 e16 37,42 34,07 9,195 Contrôle Gamme StandardH05 e17 37,32 32,73 9,950A06 e17 38,47 35,11 4,058 Well Content C(t) I.C. C(t) SQB06 e18 40,81 37,79 0,645 A01 QS1 36,02 33,04 19,00C06 e18 38,08 34,36 5,515 B01 QS1 36,06 33,22 19,00D06 e19 37,78 34,27 6,964 C01 QS2 34,01 35,6 390,00E06 e19 37,9 34,15 6,307 D01 QS2 42,26 N/A 390,00F06 e20 38,16 34,63 5,147 E01 QS3 29,99 34,11 3900,00G06 e20 39,2 34,48 2,280 F01 QS3 29,72 33,4 3900,00H06 e21 37,28 33,31 10,260 G01 QS4 26,4 33,75 39000,00A07 e21 41,82 35,38 0,293 H01 QS4 26,37 33,98 39000,00B07 e22 39,31 35,69 2,096C07 e22 37,61 33,71 7,956D07 e23 37,95 34,28 6,091 Contrôle négatifE07 e23 38,17 34,39 5,128F07 e24 38,14 34,38 5,261 Content C(t) I.C. C(t) SQG07 e24 37,92 34,65 6,214 G09 Neg Ctrl N/A 34,62 N/AH07 e25 37,33 33,41 9,875 H09 Neg Ctrl N/A 34,39 N/AA08 e25 37,47 33,94 8,870B08 e26 37,84 34,55 6,629C08 e26 37,97 34,48 5,997D08 e27 38,04 34,37 5,654E08 e27 37,53 34,6 8,487F08 e28 38,98 34,64 2,712G08 e28 38,15 34,36 5,218H08 e29 37,74 33,16 7,195A09 e29 41,52 39 0,372B09 e30 38,16 34,32 5,146C09 e30 38,22 34,72 4,908D09
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
ANNEXE F
LIMITE DE QUANTIFICATION
Limite de quantification LQ à 15UG
Gamme de calibrage QS
UG/puits Moy Log (UG/puits) C(t)
QS1 1,278753601 36,87
1,278753601 37,23
QS2 2,591064607 33,65
2,591064607 33,71
QS3 3,591064607 29,73 Pente -3,241
3,591064607 29,87 Ordonnée origine 41,514
QS4 4,591064607 26,41 Corrélation (r2) 0,992
4,591064607 26,51 Efficacité (%) 103,474
LQPCR à 15UG : 30 mesures en réplicat
Réplicat Moyenne UG/puits Moy UG/puits x' (Log) Moyenne x'LQ-1 37,55 16,7 1,223
37,87 13,4 1,124LQ-2 37,57 16,5 1,217
37,27 20,4 1,309LQ-3 37,17 22,0 1,340
37,08 23,3 1,368LQ-4 37,21 21,4 1,328
37,49 17,5 1,241LQ-5 37,53 17,0 1,229
37,63 15,8 1,198LQ-6 37,73 14,7 1,167
37,21 21,3 1,328LQ-7 36,83 27,9 1,445
37,00 24,8 1,393LQ-8 37,07 23,5 1,371
37,21 21,3 1,328LQ-9 37,73 14,8 1,167
37,68 15,2 1,183LQ-10 37,82 13,8 1,140
38,15 11,0 1,038LQ-11 37,25 20,7 1,315
37,09 23,1 1,365LQ-12 36,54 34,4 1,534
37,06 23,7 1,374LQ-13 37,35 19,3 1,285
37,26 20,5 1,312LQ-14 37,56 16,7 1,220
37,57 16,4 1,217LQ-15 37,50 17,3 1,238
37,04 24,0 1,380LQ-16 36,48 35,7 1,553
37,37 19,0 1,278LQ-17 37,13 22,5 1,352
37,43 18,3 1,260LQ-18 37,39 18,7 1,272
37,65 15,6 1,192LQ-19 36,69 30,8 1,488
37,00 24,7 1,393LQ-20 36,57 33,5 1,525
37,13 22,5 1,352LQ-21 37,70 15,0 1,177
37,18 21,8 1,337LQ-22 37,75 14,5 1,161
37,68 15,2 1,183LQ-23 37,29 20,2 1,303
37,25 20,7 1,315LQ-24 37,00 24,7 1,393
36,94 25,8 1,411LQ-25 37,29 20,1 1,303
37,62 15,9 1,201LQ-26 37,55 16,7 1,223
36,63 32,1 1,507LQ-27 36,54 34,3 1,534
37,02 24,4 1,386LQ-28 36,83 28,0 1,445
36,99 24,9 1,396LQ-29 37,58 16,4 1,214
37,15 22,3 1,346LQ-30 37,12 22,8 1,356
37,34 19,5 1,288
1,309
0,097
0,030
0,101
0,207
37,37
37,23
37,46
37,09
36,78
36,91
37,44
37,72
37,27
36,97
37,28
37,52
36,85
36,85
37,31
37,57
37,27
36,93
37,7
37,99
37,17
36,8
37,58
37,47
36,91
37,14
37,71
37,42
37,13
37,35
C(t) UG/puits
Biais
Incertitude ULQ
Ecart-type s
Moyenne x'
Exactitude de LQ ELQ
1,50E+01
1,84E+01
2,26E+01
2,63E+01
1,94E+01
1,64E+01
2,19E+01
1,50E+01
1,24E+01
1,80E+01
1,99E+01
2,80E+01
2,77E+01
1,72E+01
2,04E+01
2,73E+01
2,07E+01
1,66E+01
1,49E+01
2,44E+01
2,04E+01
2,52E+01
2,11E+01
2,94E+01
2,64E+01
1,285
1,214
1,248
1,93E+01
1,80E+01
1,84E+01
2,91E+01
1,173
1,263
1,354
1,419
1,349
1,175
1,089
2,24E+01
39000
3900
390
19
1,340
1,454
1,298
1,218
1,309
1,416
1,306
1,232
1,440
1,439
1,257
1,172
1,309
1,402
1,252
1,365
1,460
1,420
1,280
1,322
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
ANNEXE G
RENDEMENT ET ROBUSTESSE
Ren
dem
ent e
t rob
uste
sse
Rob
uste
sse
Eau
Cha
ude
San
itaire
Prot
ocol
e A
quad
ien
Prot
ocol
e A
quad
ien
W2
Niv
eau
N1
100
000
UG
/LU
G/p
uits
A (l
og)
C(t)
UG
/pui
tsM
oyen
ne U
G/p
uits
B (l
og)
log
%U
G/p
uits
A (l
og)
C(t)
UG
/pui
tsM
oyen
ne U
G/p
uits
B (l
og)
log
%E
C1N
16,
55E
+02
5,02
29,4
31,
31E
+03
EC
2N1W
2,32
E+0
46,
5725
,79
3,27
E+0
429
,67
1,10
E+0
325
,69
3,53
E+0
4E
C2N
16,
55E
+02
5,02
29,7
1,08
E+0
3E
C2N
1W2,
32E
+04
6,57
25,8
83,
03E
+04
29,7
31,
06E
+03
25,9
72,
83E
+04
EC
3N1
6,28
E+0
25,
0029
,41
1,19
E+0
3E
C3N
1W4,
31E
+02
4,84
30,3
6,48
E+0
229
,35
1,25
E+0
330
,32
6,40
E+0
2E
C4N
16,
28E
+02
5,00
29,3
21,
27E
+03
EC
4N1W
4,31
E+0
24,
8430
,16
7,22
E+0
229
,31
1,29
E+0
330
,22
6,89
E+0
2E
C5N
14,
31E
+02
4,84
29,2
41,
46E
+03
E5N
1W-1
003,
27E
+04
6,72
30,5
37,
23E
+02
29,5
1,20
E+0
330
,57
7,02
E+0
2E
C6N
14,
31E
+02
4,84
29,3
11,
39E
+03
E6N
1W-1
003,
27E
+04
6,72
30,4
8,00
E+0
229
,38
1,31
E+0
330
,36
8,29
E+0
2E
7N1-
100
3,27
E+0
46,
7231
5,01
E+0
2E
7N1W
-100
3,27
E+0
46,
7230
,38
8,17
E+0
230
,59
6,91
E+0
230
,31
8,58
E+0
2E
8N1-
100
3,27
E+0
46,
7230
,88
5,52
E+0
2E
C8N
1W7,
10E
+03
6,06
26,8
79,
45E
+03
30,6
26,
78E
+02
26,7
41,
04E
+04
E9N
1-10
03,
27E
+04
6,72
30,8
5,89
E+0
2E
9N1W
7,00
E+0
47,
0523
,82
9,00
E+0
430
,76,
32E
+02
23,8
98,
51E
+04
EC
10N
17,
10E
+03
6,06
25,8
2,11
E+0
4E
10N
1W7,
00E
+04
7,05
23,5
71,
08E
+05
25,6
62,
34E
+04
23,6
31,
03E
+05
Ren
dem
ent m
oyen
pou
r le
nive
au 1
00 0
00 U
G/L
-0,3
941
%R
ende
men
t moy
en p
our l
e ni
veau
100
000
UG
/L-0
,45
36%
Niv
eau
N2
1 00
0 U
G/L
UG
/pui
tsA
(log
)C
(t)U
G/p
uits
Moy
enne
UG
/pui
tsB
(log
)lo
g%
UG
/pui
tsA
(log
)C
(t)U
G/p
uits
Moy
enne
UG
/pui
tsB
(log
)lo
g%
EC
1N2
2,32
E+0
46,
5730
,25
9,11
E+0
2E
C1N
2W6,
55E
+02
5,02
34,5
3,21
E+0
130
,23
9,23
E+0
235
,31,
78E
+01
EC
2N2
2,32
E+0
46,
5730
,13
1,00
E+0
3E
C2N
2W6,
55E
+02
5,02
34,7
42,
68E
+01
30,1
21,
01E
+03
35,0
12,
20E
+01
EC
3N2
4,04
E+0
46,
8130
,24
8,24
E+0
2E
C3N
2W6,
28E
+02
5,00
34,9
72,
10E
+01
30,0
59,
53E
+02
36,8
5,51
E+0
0E
C4N
24,
04E
+04
6,81
30,0
89,
29E
+02
EC
4N2W
6,28
E+0
25,
0034
,74
2,46
E+0
129
,96
1,02
E+0
335
,97
1,01
E+0
1E
C5N
24,
04E
+04
6,81
30,1
29,
00E
+02
EC
5N2W
2,32
E+0
46,
5731
,16
4,39
E+0
229
,98
1,01
E+0
331
,05
4,79
E+0
2E
C6N
23,
27E
+04
6,72
30,2
98,
76E
+02
EC
6N2W
2,32
E+0
46,
5730
,97
5,09
E+0
230
,21
9,28
E+0
231
,21
4,19
E+0
2E
C7N
23,
27E
+04
6,72
30,2
49,
07E
+02
EC
7N2W
4,31
E+0
24,
8434
,78
2,09
E+0
130
,11,
01E
+03
35,5
21,
18E
+01
EC
8N2
7,10
E+0
36,
0631
,82
2,28
E+0
2E
C8N
2W4,
31E
+02
4,84
34,8
2,07
E+0
132
,06
1,91
E+0
235
,57
1,15
E+0
1E
C9N
27,
21E
+04
7,06
30,0
51,
07E
+03
EC
9N2W
7,10
E+0
36,
0632
,14
1,80
E+0
230
,09
1,04
E+0
331
,91
2,14
E+0
2E
C10
N2
7,21
E+0
47,
0630
,38
8,41
E+0
2E
10N
2W7,
00E
+04
7,05
30,0
58,
90E
+02
30,1
59,
95E
+02
29,8
71,
02E
+03
Ren
dem
ent m
oyen
pou
r le
nive
au 1
000
UG
/L-0
,29
51%
Ren
dem
ent m
oyen
pou
r le
nive
au 1
000
UG
/L-0
,16
69%
Ren
dem
ent m
oyen
Eau
cha
ude
sani
taire
Aqu
adie
n-0
,34
46%
Ren
dem
ent m
oyen
Eau
cha
ude
sani
taire
Aqu
adie
n W
2-0
,30
50%
1,07
E+0
34,
53-0
,49
33%
4,59
-0,4
437
%-0
,48
3,40
E+0
46,
0933
%1,
20E
+03
1,22
E+0
34,
59-0
,41
39%
1,28
E+0
34,
61
3,21
E+0
15,
060,
0411
0%
-0,3
941
%
2,43
E+0
14,
94
88%
-0,0
883
%
2,93
E+0
46,
02-0
,55
28%
9,17
E+0
26,
47-0
,10
79%
1,00
E+0
36,
51-0
,06
87%
4,88
4,58
E+0
26,
22-0
,35
45%
4,62
E+0
26,
22-0
,35
45%
1,32
E+0
34,
63-0
,21
61%
-0,2
063
%
6,44
E+0
24,
37-0
,47
34%
7,05
E+0
24,
40-0
,43
37%
2,09
E+0
14,
880,
0410
9%
2,07
E+0
14,
870,
0310
8%
-0,1
275
%2,
10E
+01
-0,0
52,
46E
+01
34%
8,86
E+0
26,
45-0
,28
53%
4,95
9,74
E+0
26,
49-0
,23
58%
9,54
E+0
26,
48-0
,24
57%
5,89
E+0
26,
27-0
,52
30%
1,35
E+0
34,
648,
14E
+02
6,47
-0,3
347
%
6,12
E+0
26,
29-0
,50
31%
6,10
E+0
26,
29-0
,50
31%
7,12
E+0
26,
41-0
,38
41%
8,37
E+0
26,
48-0
,31
48%
9,02
E+0
26,
46-0
,33
46%
9,58
E+0
26,
49-0
,31
49%
1,06
E+0
5
8,75
E+0
46,
50
6,58
0475
6
-0,5
5
-0,4
7
28%
2,22
E+0
45,
85-0
,20
63%
9,92
E+0
35,
55-0
,50
31%
2,09
E+0
25,
82-0
,23
59%
62%
1,05
E+0
36,
53-0
,53
29%
9,54
E+0
26,
48-0
,56
27%
1,96
E+0
25,
85-0
,21
9,15
E+0
26,
47-0
,60
25%
Ech
antil
lon
Val
eur d
u do
page
Rés
ulta
t ana
lyse
Ren
dem
ent
Ech
antil
lon
Val
eur d
u do
page
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IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
Ren
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Aér
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Prot
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Prot
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W2
Niv
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100
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UG
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G/p
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G/p
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A (l
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C(t)
UG
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%T1
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2,32
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46,
5725
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4,01
E+0
4T1
N1W
6,55
E+0
25,
0229
,71
1,08
E+0
325
,35
4,64
E+0
429
,72
1,07
E+0
3T2
N1
2,32
E+0
46,
5725
,39
4,52
E+0
4T2
N1W
2,32
E+0
46,
5726
,11
2,52
E+0
425
,26
5,00
E+0
426
,12,
55E
+04
T3N
14,
04E
+04
6,81
23,7
1,45
E+0
5T3
N1W
4,31
E+0
24,
8430
,35
6,26
E+0
223
,57
1,61
E+0
530
,41
5,95
E+0
2T4
N1
4,04
E+0
46,
8123
,64
1,52
E+0
5T4
N1W
4,31
E+0
24,
8430
,26
6,68
E+0
223
,58
1,59
E+0
530
,33
6,33
E+0
2T5
N1
4,04
E+0
46,
8123
,61,
57E
+05
T5N
1W4,
04E
+04
6,81
24,9
65,
35E
+04
23,3
1,98
E+0
525
5,17
E+0
4T6
N1-
100
3,27
E+0
46,
7231
,02
4,95
E+0
2T6
N1W
4,04
E+0
46,
8125
,24
4,27
E+0
430
,81
5,80
E+0
225
,03
5,04
E+0
4T7
N1-
100
3,27
E+0
46,
7231
,06
4,78
E+0
2T7
N1W
4,04
E+0
46,
8125
,16
4,57
E+0
430
,78
5,95
E+0
225
,06
4,95
E+0
4T8
N1-
100
3,27
E+0
46,
7231
,09
4,68
E+0
2T8
N1W
-100
3,27
E+0
46,
7231
,47
3,47
E+0
230
,79
5,93
E+0
231
,32
3,90
E+0
2T9
N1
7,10
E+0
36,
0626
,26
1,49
E+0
4T9
N1W
7,10
E+0
36,
0626
,14
1,63
E+0
426
,11,
68E
+04
25,9
21,
92E
+04
T10N
17,
21E
+04
7,06
23,4
11,
46E
+05
T10N
1W7,
00E
+04
7,05
23,3
1,32
E+0
523
,45
1,42
E+0
523
,41
1,22
E+0
5
Ren
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G/L
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149
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100
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UG
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Niv
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G/L
UG
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(t)U
G/p
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Moy
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UG
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UG
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(log
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(t)U
G/p
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Moy
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UG
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T1N
26,
55E
+02
5,02
34,7
22,
72E
+01
T1N
2W2,
32E
+04
6,57
31,5
33,
24E
+02
34,7
72,
62E
+01
31,8
2,62
E+0
2T2
N2
6,55
E+0
25,
0234
,58
3,01
E+0
1T2
N2W
2,32
E+0
46,
5731
,71
2,81
E+0
234
,72,
77E
+01
31,5
33,
24E
+02
T3N
26,
28E
+02
5,00
34,3
63,
27E
+01
T3N
2W4,
31E
+02
4,84
34,8
2,06
E+0
134
,39
3,18
E+0
135
,64
1,08
E+0
1T4
N2
6,28
E+0
25,
0034
,94
2,13
E+0
1T4
N2W
4,04
E+0
46,
8130
,74
5,51
E+0
234
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3,04
E+0
130
,98
4,58
E+0
2T5
N2
2,32
E+0
46,
5730
,86
5,56
E+0
2T5
N2W
4,04
E+0
46,
8130
,85,
27E
+02
30,7
85,
91E
+02
30,9
74,
60E
+02
T6N
22,
32E
+04
6,57
30,8
45,
64E
+02
T6N
2W4,
04E
+04
6,81
30,9
4,86
E+0
230
,73
6,18
E+0
230
,95
4,69
E+0
2T7
N2
4,04
E+0
46,
8129
,41,
59E
+03
T7N
2W7,
10E
+03
6,06
31,7
62,
39E
+02
29,3
71,
64E
+03
32,0
31,
95E
+02
T8N
24,
04E
+04
6,81
29,4
31,
56E
+03
T8N
2W7,
21E
+04
7,06
30,4
57,
96E
+02
29,7
81,
18E
+03
30,4
8,28
E+0
2T9
N2-
100
3,27
E+0
46,
72N
/AN
/AT9
N2W
7,00
E+0
47,
0530
,29
7,46
E+0
235
,35
1,66
E+0
130
,01
9,17
E+0
2T1
0N2
7,10
E+0
36,
0631
,45
3,01
E+0
2T1
0N2W
7,00
E+0
47,
0530
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9,18
E+0
231
,35
3,24
E+0
229
,68
1,18
E+0
3
Ren
dem
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r le
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au 1
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UG
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Ren
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r le
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UG
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Ren
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Tou
r aér
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Aqu
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Ren
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Tou
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Aqu
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4,31
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46,
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1,07
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34,
59-0
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2,67
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14,
93-0
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37%
82%
2,54
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45,
96-0
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25%
2,91
E+0
2
2,89
E+0
14,
97-0
,05
88%
6,10
E+0
24,
34-0
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5,02
E+0
2
2,06
E+0
110
8%
6,21
-0,5
230
%
4,87
81%
3,22
E+0
15,
010,
0110
3%
4,75
E+0
46,
18-0
,39
41%
5,73
E+0
26,
26-0
,31
5,90
E+0
26,
28-0
,29
51%
2,55
E+0
14,
91-0
,09
29%
50%
29%
6,02
-0,5
528
%
3,02
E+0
26,
04-0
,53
31%
32%
32%
5,36
E+0
26,
23
32%
1,53
E+0
56,
69-0
,04
91%
6,50
E+0
24,
37-0
,47
34%
0,03
1,55
E+0
56,
70-0
,03
93%
35%
1,76
E+0
56,
750,
0210
5%5,
26E
+04
6,28
-0,4
5
4,64
E+0
46,
22-0
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4,75
E+0
46,
23-0
,50
-0,4
9
5,34
E+0
26,
23-0
,50
5,27
E+0
26,
23-0
,50
32%
31%
1,66
E+0
1
1,36
E+0
3
1,61
E+0
3
3,12
E+0
26,
00-0
,06
1,58
E+0
45,
70-0
,35
88%
1,77
E+0
45,
80-0
,25
56%
45%
2,16
E+0
25,
89-0
,16
6,73
0,00
99%
-0,0
491
%6,
69
1,44
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56,
66
68%
6,76
0,04
108%
4,77
E+0
2
-0,4
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%
3,68
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26,
12-0
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1,27
E+0
56,
66-0
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1,04
E+0
36,
57-0
,48
33%
8,27
E+0
26,
47-0
,58
27%
25%
29%
4,93
E+0
26,
20-0
,53
6,18
-0,5
4
8,12
E+0
26,
47-0
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Ech
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Rés
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Ren
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Val
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Ech
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Val
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Rés
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Rés
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Ech
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Rés
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IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
Ren
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Eau
Min
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Prot
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Prot
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Niv
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UG
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G/p
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%M
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6,55
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25,
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1,15
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1N1W
6,55
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25,
0230
,16
7,73
E+0
229
,65
1,12
E+0
330
,22
7,37
E+0
2M
2N1
6,55
E+0
25,
0229
,37
1,37
E+0
3M
2N1W
6,55
E+0
25,
0229
,64
1,13
E+0
329
,47
1,28
E+0
330
,04
8,40
E+0
2M
3N1
6,28
E+0
25,
0029
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1,22
E+0
3M
3N1W
6,28
E+0
25,
0029
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8,42
E+0
229
,22
1,37
E+0
329
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9,60
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2M
4N1
6,28
E+0
25,
0029
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1,30
E+0
3M
4N1W
6,28
E+0
25,
0029
,74
9,43
E+0
229
,14
1,46
E+0
329
,82
8,89
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2M
5N1
2,32
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46,
5725
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3,48
E+0
4M
5N1W
4,31
E+0
24,
8430
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8,09
E+0
225
,65
3,65
E+0
430
,04
7,92
E+0
2M
6N1
2,32
E+0
46,
5725
,74
3,41
E+0
4M
6N1W
4,31
E+0
24,
8429
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8,24
E+0
225
,64
3,70
E+0
430
8,16
E+0
2M
7N1
4,31
E+0
24,
8429
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1,12
E+0
3M
7N1W
-100
3,27
E+0
46,
7229
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1,92
E+0
329
,65
1,07
E+0
330
,83
5,73
E+0
2M
8N1
4,31
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24,
8429
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1,37
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3M
8N1W
-100
3,27
E+0
46,
7230
,76
6,08
E+0
229
,54
1,17
E+0
330
,69
6,39
E+0
2M
9N1
4,04
E+0
46,
8125
,02
5,11
E+0
4M
9N1W
7,10
E+0
36,
0626
,68
1,09
E+0
424
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5,76
E+0
426
,59
1,16
E+0
4M
10N
14,
04E
+04
6,81
25,1
74,
52E
+04
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N1W
7,21
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47,
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75,
73E
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76,
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Ren
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G/L
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Niv
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Moy
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Moy
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M1N
26,
55E
+02
5,02
33,6
65,
91E
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M1N
2W2,
32E
+04
6,57
31,8
42,
53E
+02
34,6
62,
84E
+01
31,7
42,
75E
+02
M2N
26,
55E
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5,02
34,2
3,99
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2N2W
2,32
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5731
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2,66
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234
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2,72
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131
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2,56
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25,
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4,08
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7,10
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36,
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+02
33,8
84,
62E
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32,9
39,
90E
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22,
32E
+04
6,57
31,0
94,
64E
+02
31,0
84,
66E
+02
M5N
22,
32E
+04
6,57
31,1
34,
49E
+02
31,0
14,
94E
+02
M6N
24,
04E
+04
6,81
29,8
51,
12E
+03
29,8
21,
15E
+03
M7N
24,
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+04
6,81
29,8
11,
15E
+03
29,8
51,
12E
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M8N
27,
10E
+03
6,06
32,5
31,
34E
+02
32,6
11,
26E
+02
M9N
27,
21E
+04
7,06
30,2
29,
47E
+02
30,1
99,
65E
+02
M10
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7,00
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47,
0530
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230
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8,71
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2
Ren
dem
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pou
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au 1
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Ren
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Ren
dem
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Ren
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43-0
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4,63
1,33
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3
1,14
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56
32%
1,38
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64-0
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51-0
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429
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41%
4,48
3,56
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1,29
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62-0
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41%
4,34
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15,
140,
14
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6,18
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4
51%
4,54
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9
6,06
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131
%
31%
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52-0
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33%
4,47
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6,35
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436
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Ren
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Rés
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5,09
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21-0
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1,13
E+0
36,
56-0
,17
68%
1,14
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36,
56-0
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61%
6,23
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2
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5,61
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1,08
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2,61
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138%
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45,
621,
30E
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06,
174,
65E
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3,29
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15,
020,
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6,44
9,56
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26,
49
8,59
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Rés
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Val
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Val
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Ech
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Ren
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Ech
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Rés
ulta
t ana
lyse
Ren
dem
ent
Val
eur d
u do
page
Ech
antil
lon
Rés
ulta
t ana
lyse
Val
eur d
u do
page
6,85
E+0
46,
39-0
,60
25%
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
ANNEXE H
SELECTIVITE
Sél
ectiv
itéSo
uche
s ci
bles
: Le
gion
ella
spp
.
Taux
cib
le
inoc
ulum
(Eq
UG
/pui
ts)
Ct (
moy
)U
G/p
uits
Dét
ectio
n Le
gion
ella
1L.
pne
umop
hila
ser
1C
IP 1
0385
4T1,
00E
+03
33,2
110
5D
étec
té
2L.
pne
umop
hila
ser
2C
HU
L LG
100
7 30
021,
00E
+03
33,0
012
2D
étec
té
3L.
pne
umop
hila
ser
3C
HU
L LG
101
6 20
141,
00E
+03
33,1
910
9D
étec
té
4L.
pne
umop
hila
ser
4C
HU
L LG
100
6 30
101,
00E
+03
33,0
311
9D
étec
té
5L.
pne
umop
hila
ser
5C
HU
L LG
100
8 50
131,
00E
+03
33,1
910
6D
étec
té
6L.
pne
umop
hila
ser
6A
TCC
332
151,
00E
+03
32,9
810
1D
étec
té
7L.
pne
umop
hila
ser
7C
HU
L LG
102
2 11
051,
00E
+03
32,9
312
9D
étec
té
8L.
pne
umop
hila
ser
8C
HU
L LG
100
9 30
091,
00E
+03
33,0
012
3D
étec
té
9L.
pne
umop
hila
ser
9C
HU
L LG
092
5 40
121,
00E
+03
32,9
213
0D
étec
té
10L.
pne
umop
hila
ser
10
CH
UL
LG 1
009
2018
1,00
E+0
332
,63
162
Dét
ecté
11L.
pne
umop
hila
ser
11
CH
UL
LG 0
841
3021
1,00
E+0
332
,58
168
Dét
ecté
12L.
pne
umop
hila
ser
12
CH
UL
LG 1
009
3041
1,00
E+0
332
,54
174
Dét
ecté
13L.
pne
umop
hila
ser
13
CH
UL
LG 1
022
1006
1,00
E+0
332
,88
133
Dét
ecté
14L.
pne
umop
hila
ser
14
CH
UL
LG 0
916
4027
1,00
E+0
332
,57
168
Dét
ecté
15L.
pne
umop
hila
ser
15
CH
UL
LG 0
312
4049
1,00
E+0
332
,61
163
Dét
ecté
16Le
gion
ella
ani
saC
IP 1
0387
01,
00E
+03
32,2
117
7D
étec
té
17Le
gion
ella
birm
ingh
amsi
sC
HU
L H
L 06
2840
371,
00E
+03
32,2
117
7D
étec
té
18Le
gion
ella
boz
eman
iiC
IP 1
0387
2 (e
q A
TCC
332
17)a
1,00
E+0
332
,66
127
Dét
ecté
19Le
gion
ella
che
rrii
CH
UL
HL
0521
4024
1,00
E+0
332
,30
166
Dét
ecté
20Le
gion
ella
cin
cinn
atie
nsis
CIP
103
875
1,00
E+0
332
,09
193
Dét
ecté
21Le
gion
ella
dun
mof
iiC
IP 1
0387
6 (é
q A
TCC
332
79)b
1,00
E+0
332
,26
170
Dét
ecté
22Le
gion
ella
ery
thra
CH
UL
LG07
1301
21,
00E
+03
32,6
712
6D
étec
té
23Le
gion
ella
feel
eii
CH
UL
LG07
5030
221,
00E
+03
32,2
816
9D
étec
té
24Le
gion
ella
gor
man
iiC
HU
L LG
102
3200
71,
00E
+03
32,6
313
3D
étec
té
25Le
gion
ella
hac
kelia
eC
IP10
3844
1,00
E+0
332
,50
143
Dét
ecté
26Le
gion
ella
jord
anis
CH
UL
LG 0
9455
020
1,00
E+0
332
,25
172
Dét
ecté
27Le
gion
ella
lans
inge
nsis
ATC
C 4
3751
1,00
E+0
332
,15
185
Dét
ecté
28Le
gion
ella
long
beac
hae
CH
UL
HL
0638
3034
1,00
E+0
332
,56
137
Dét
ecté
29Le
gion
ella
mac
each
erni
iC
HU
L LG
092
2100
91,
00E
+03
32,2
117
7D
étec
té
30Le
gion
ella
mic
dade
iC
IP 1
0388
2 (é
q A
TCC
332
18)c
1,00
E+0
332
,31
165
Dét
ecté
31Le
gion
ella
oak
ridge
nsis
CH
UL
LG 0
7122
004
1,00
E+0
332
,60
133
Dét
ecté
32Le
gion
ella
par
isie
nsis
CH
UL
LG 0
8513
015
1,00
E+0
332
,19
180
Dét
ecté
33Le
gion
ella
sai
nthe
lens
iC
HU
L H
L 06
3530
041,
00E
+03
32,2
017
9D
étec
té
34Le
gion
ella
tucs
onen
sis
CH
UL
LG 0
8495
014
1,00
E+0
332
,37
158
Dét
ecté
35Le
gion
ella
wad
swor
thii
CIP
103
886
1,00
E+0
332
,09
193
Dét
ecté
a Fluo
ribac
ter b
ozem
anae
b Fluo
ribac
ter d
umof
iic Ta
tlock
ia m
icda
deiSo
uche
Orig
ine
IQ C
heck
Leg
ione
lla s
pp
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01
Sél
ectiv
itéEx
clus
ivité
Taux
cib
le
inoc
ulum
(Eq
UG
/pui
ts)
Ct (
moy
)U
G/p
uits
Dét
ectio
n
1A
erom
onas
hyd
roph
ilaE
nviro
nnem
ent
1,00
E+0
4N
/A-
Non
dét
ecté
2A
lcal
igen
es fa
ecal
isC
IP 6
0.80
1,00
E+0
4N
/A-
Non
dét
ecté
3B
acill
us s
ubtil
isC
CM
199
91,
00E
+04
N/A
-N
on d
étec
té
4B
urkh
olde
ria c
epac
iaE
au d
e do
uche
, La
chap
elle
St M
esni
n1,
00E
+04
N/A
-N
on d
étec
té
5C
lost
ridiu
m
Eau
de
puits
, Bog
hni
1,00
E+0
4N
/A-
Non
dét
ecté
6E
nter
obac
ter a
erog
enes
Env
ironn
emen
t1,
00E
+04
495,
17E
-04
Non
dét
ecté
7E
sche
richi
a co
liE
au d
’alim
enta
tion,
Lie
ncou
rt1,
00E
+04
N/A
-N
on d
étec
té
8Fl
avob
acte
rium
alg
icol
aE
nviro
nnem
ent
1,00
E+0
4N
/A-
Non
dét
ecté
9K
lebs
iella
oxy
toca
ATC
C 4
9473
1,00
E+0
445
,42
7,56
E-0
3N
on d
étec
té
10Li
ster
ia m
onoc
ytog
enes
CC
M 5
576
1,00
E+0
4N
/A-
Non
dét
ecté
11P
rote
us v
ulga
risE
nviro
nnem
ent
1,00
E+0
443
,55
3,09
E-0
2N
on d
étec
té
12P
seud
omon
as a
erug
inos
aE
au d
’alim
enta
tion,
Lill
e1,
00E
+04
42,7
5,83
E-0
2N
on d
étec
té
13P
seud
omon
as fl
uore
scen
sE
nviro
nnem
ent
1,00
E+0
4N
/A-
Non
dét
ecté
14P
seud
omon
as p
utid
aE
nviro
nnem
ent
1,00
E+0
4N
/A-
Non
dét
ecté
15S
erra
tia m
arce
scen
sE
nviro
nnem
ent
1,00
E+0
443
,13
4,21
E-0
2N
on d
étec
té
16S
teno
troph
omon
as m
alto
phila
Can
al d
e la
Deû
le, L
ille
1,00
E+0
4N
/A-
Non
dét
ecté
Orig
ine
Souc
heIQ
Che
ck L
egio
nella
spp
IPL santé, environnement durables Nord – 1 rue du Professeur Calmette – 59046 LILLE cedex
iQ-Check™ Quanti Legionella spp. - Ext 2011 - v01