eV on Molecular EQA for Measles and Rubella Process and results of 1 st molecular PT exercise in EUR Oliver Donoso Mantke 1 and Heinz Zeichhardt 123 1 INSTAND ID: 934173
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Slide1
1
New Collaboration between WHO and INSTAND e.V. on Molecular EQA for Measles and Rubella Process and results of 1st molecular PT exercise in EUR
Oliver Donoso Mantke
1
and
Heinz
Zeichhardt
1,2,3
1
INSTAND
e.V. - Gesellschaft zur Förderung der Qualitätssicherung in medizinischen Laboratorien e.V.,
Düsseldorf
2
Professor
of
Virology
(
retired
)
Charité
Universitätsmedizin Berlin, Campus Benjamin Franklin, Institut für Virologie,
Berlin
3
Institut für Qualitätssicherung in der Virusdiagnostik - IQVD, Berlin
at GBD Gesellschaft für Biotechnologische Diagnostik mbH, Berlin
Vanessa
Lindig
3
and
Hans-Peter Grunert
4
3
Institut für Qualitätssicherung in der Virusdiagnostik - IQVD, Berlin
at GBD Gesellschaft für Biotechnologische Diagnostik mbH, Berlin
4
GBD Gesellschaft
für Biotechnologische Diagnostik
mbH
, Berlin
in
cooperation
with
Sabine Santibanez
and
Annette Mankertz
Nationales Referenzzentrum für Masern, Mumps und Röteln
am Robert Koch-Institut, Berlin
FG 12 Masern, Mumps, Röteln und Viren bei Abwehrschwäche
Slide2International14 Organistions / Scientific SocietiesNational28 Organistions / Scientific Societies
Slide3Slide4INSTAND EQA Schemes
Slide5O. Adams Universitätsklinikum DüsseldorfA. Baillot Niedersächsisches Landesgesundheitsamt, HannoverS. Becker / M. Eickmann Philipps Universität Marburg, Nat. Konsiliarlab. FilovirenC.-T. Bock / S. Niendorf / A. Mas-Marques Robert Koch-Institut, Berlin; Nat. Konsiliarlab. Noroviren
; Nat. Konsiliarlab. RotavirenS. Diedrich Robert Koch-Institut, Berlin; NRZ Poliomyelitis und Enteroviren Regionales Referenzlabor der WHO/EURO für PoliomyelitisU. Dittmer / S. Ross / M. Roggendorf Universitätsklinik Essen; NRZ HCV; Nat. Konsiliarlab. TollwutG. Dobler Institut für Mikrobiologie der Bundeswehr, München; Nat. Konsiliarlab. FSMECh. Drosten / A.-M. Eis-Hübinger Universität Bonn; Nat. Konsiliarlabor für
Coronaviren
J.
Eberle
/ L.
Gürtler
Ludwig-
Maximillians
-
Universität München, Max-von-Pettenkofer InstitutG. Enders / M. Enders Labor Enders, StuttgartJ. Fuhrmann MVZ Labor 28 GmbH, BerlinA. Gessner / B. Schmidt / J. Wenzel Universität Regensburg; Nat. Konsiliarlab. HAV und HEVS. Günther / J. Schmidt-Chanasit / P. Emmerich Bernhard-Nocht-Institut, Hamburg; NRZ trop. Infektionserreger, WHO CCG. Harms-Zwingenberger Institut für Tropenmedizin, BerlinH.H. Hirsch Universität Basel, Dept. BiomedizinD. Huzly / M. Panning Universitätsklinikum FreiburgG. Jahn / K. Hamprecht Universitätsklinikum Tübingen; Nat. Konsiliarlab. CMVA. Karl / K. Frank / K. Gubbe DRK-Blutspendedienst Ost, PlauenV. Kempf / H. Rabenau / A. Berger / M. Stürmer Universitätsklinikum Frankfurt; NRZ RetrovirenD. Krüger / J. Hofmann Charité Universitätsmedizin Berlin, CCM; Nat. Konsiliarlab.Hantaviren; Labor Berlin – Charité Vivantes GmbHC. Kücherer Robert Koch-Institut, BerlinU.G. Liebert Universität LeipzigA. Mankertz / S. Santibanez Robert Koch-Institut, Berlin; NRZ Masern, Mumps und RötelnT. Mertens / D. Michel Universitätsklinikum Ulm; Nat. Konsiliarlab.CMVS. Modrow Universität Regensburg; Nat. Konsiliarlab. ParvovirenS. Nick / H. Scheiblauer Paul-Ehrlich-Institut, Langen; Prüflabor für IVDM. Nübling / M. Chudy / S.A. Baylis / J. Kreß Paul-Ehrlich-Institut, Langen, WHO CCH. Pfister / U. Wieland / S. Silling / R. Kaiser Uniklinik Köln; NRZ Papillom- und PolyomavirenB. Weißbrich Universität WürzburgA. Sauerbrei / P. Wutzler Universitätsklinikum Jena; Nat. Konsiliarlab. HSV und VZVT. Schulz / A. Heim Medizinische Hochschule Hannover; Nat. Konsiliarlab. AdenovirenT. Schulz / W. Puppe / C. Schmitt Medizinische Hochschule Hannover; Nat. Konsiliarlab. EBV, HHV 6, 7, 8B. Schweiger Robert Koch-Institut, Berlin; NRZ Influenza; Nat. Konsiliarlab. RSV, Parainfluenzaviren, HMPVS. Smola, N. Müller-Lantzsch, J. Rissland Universitätsklinikum des SaarlandesK. Überla / K. Korn Universitätsklinikum ErlangenJ. Ziebuhr, D. Glebe / C. Schüttler / W. Gerlich Universität Gießen; NRZ HBV und HDV
INSTAND Target Value Laboratories
(in total: 36;
6
for
MeV+RubV
)
Slide6INSTAND EQA Schemes (68 Schemes)in Virus Immunology and Genome Detection 2016Serology and Antigen Detection
Virus Genome Detection and TypingHIV-1/2Herpes simplex Viruses
HIV-1 (RNA)
HIV-2 (RNA)
Measles Rubella
Mumps
Viruses
HIV-1 p24 Ag
Varicella Zoster VirusHIV-1 Resistance + TropismAdenovirusesHTLV-1/2Epstein Barr VirusHepatitis A VirusNorovirus RotavirusHepatitis A VirusResp. Sync. Virus AgHepatitis B Virus + Genotyping + ResistanceCoronavirusHepatitis B Virus, Prg. IInfluenza A and B Ag +A/H1N1 pdm 2009+A/H5N1+A/H7N9Hepatitis C Virus+ Genotyping + Resistance
Enteroviruses + Enterovirus (WHO/RKI)
Hepatitis B Virus, Prg. II
Rubella VirusMeasles VirusMumps VirusHepatitis D VirusHepatitis E VirusHuman RhinovirusesResp. Syncytial VirusHum. MetapneumovirusParainfluenza virusesHepatitis C VirusTBE VirusCytomegalovirus+ ResistanceInfluenza A and B Ag +A/H1N1 pdm 2009+A/H5N1+A/H7N9Hepatitis D VirusHantavirusEpstein-Barr VirusBK VirusJC VirusHepatitis E VirusDengue VirusNEW: Chikungunya VirusNEW: Zika VirusHerpes simplex VirusesDengue VirusWest Nile VirusChikungunya VirusNEW: Zika VirusParvovirus B19Rabies VirusVaricella Zoster VirusHum. Papilloma VirusesCytomegalovirusBSE (PrPsc) (2002-2007)Parvovirus B19Rabies Virus
Zeichhardt – SoGAT Graz – 28.05.2015
Slide7Accreditation issues1- Rolling out 2015 molecular PT for the European regionInvite NRLs performing molecular testing in routine to enrol in Instand molecular PT (Nov 2015 round)NRLs to pay for the fees, WHO to support labs in needInstand to expand protocols to get full set of resultsContinue collaboration to harmonize Instand and CDC molec PT in coming rounds7
Slide8Summary of organizational data:No. of laboratories invited for participation: 37 (call of participation beginning Sep 2015)No. of laboratories registered for participation: 34 (as of 7 Oct 2015)Bulk shipment of sample panels to RRLs: 18.11.2015No. of laboratories reporting results: 34
(from 33 countries)(as of 27 May 2016)Response rate: 100%PCR/NAT - Measles Virus Collaboration – WHO EURO and INSTAND (991) Nov 2015
Slide9Sample properties (as discussed in Aug 2015):PCR/NAT - Measles Virus Collaboration – WHO EURO and INSTAND (991) Nov 2015 Sample No.
Sample sourceIdentical Named Strain and Distinct sequence ID in MeaNSDilution
991001
Lysate of cells infected with measles virus
(
wild-type measles
strain;
genotype
D8
)$MVs/Frankfurt Main.DEU/17.11, 2266undiluted991002Lysate of non-infected cells----991003Lysate of cells infected with measles virus (wild-type measles strain; genotype B3)$MVi/Harare.ZWE/38.09, 1310undiluted991004Lysate of cells infected with measles virus (wild-type measles strain;
genotype
D4
)$MVs/Manchester.GBR/10.09, 1306undiluted$ Infectious viruses are chemically inactivated on the sample FTA disk.
Slide10PCR/NAT - Measles Virus Collaboration – WHO EURO and INSTAND (991) Nov 2015 Summary of qualitative results and typing results:
& The success rates for all 4 samples in test categories 20 and 40, respectively, refer to the number of participating laboratories. Laboratories having reported results obtained by several methods are recorded only once.
§
One
participant (Participant No. 1191), who was registered for genotyping, did not report results in test category 40.
This
was evaluated as "false" result in test category 40.
# One
participant (Participant No. 49690), who was registered for genotyping, was not able to report results in test category 40 due to weak PCR signals. This was evaluated as "false" result in test category 40. Sample 991001Sample 991002Sample 991003Sample 991004Successratefor all samples&Sample propertiesMeasles virus positive(genotype D8) 104.0 PFU/ml on Vero/hSLAMMeasles virus negativeMeasles virus positive(genotype B3)
10
5.5
PFU/ml on Vero/hSLAMMeasles virus positive(genotype D4) 104.1 PFU/ml on Vero/hSLAMDilutionundiluted----undilutedundiluted"correct" results for qualitative genome detection (test category 20)(basis for receiving a certificate of successful participation)qual. detectionpositive100% (38/38)negative100% (38/38)positive100% (38/38)positive100% (38/38)
100%
&
(34/34)
&
"
correct
"
results
for
type
identification
(
test
category
40)
(basis for receiving a certificate of successful participation)
type
identification
D8
93.3%
(
28/30)
negative/
nd
93.3%
(
28/30)
B3
93.3%
(
28/30)
D4
93.3%
(
28/30)
93.3%
&,§,#
(28/30)
&,§,#
Slide11PCR/NAT - Measles Virus Collaboration – WHO EURO and INSTAND (991) Nov 2015 Link to results from regular INSTAND EQA scheme (386):&
& The success rates for all 4 samples in test categories 20 and 40, respectively, refer to the number of participating laboratories. Laboratories having reported results obtained by several methods are recorded only once.
Sample 991001
Sample 991002
Sample 991003
Sample 991004
Success
rate
for all samples
&
= 386015= 386014= 386016= 386013Sample propertiesMeasles virus positive(genotype D8)Measles virus negativeMeasles virus positive(genotype B3)Measles virus positive
(genotype
D4)
Dilutionundiluted----undilutedundiluted"correct" results for qualitative genome detection (test category 20)(basis for receiving a certificate of successful participation)qual. detectionpositive100% (38/38)negative100% (38/38)positive100% (38/38)positive100% (38/38)100%&(34/34)&qual. detection
positive
96.6%
(29/30)
negative
96.6%
(29/30)
positive
93.3%
(28/30)
positive
96.6%
(29/30)
93.1%
&
(27/29)
&
"
correct
"
results
for
type
identification
(
test
category
40)
(basis for receiving a certificate of successful participation)
type
identification
D8
93.3%
(
28/30)
negative/
nd
93.3%
(
28/30)
B3
93.3%
(
28/30)
D4
93.3%
(
28/30)
93.3%
&
(28/30)
&
type
identification
D8
100
%
(
2/2)
negative/
nd
100
%
(
2/2)
B3
100
%
(
2/2)
D4
100
%
(
2/2)
100%
&
(2/2)
&
Slide12New test categoryfor sequence quality:Success rate for all samples refers
to the number of participating laboratories(in 1st round/Nov 2015 not evaluated w/o consequences for certificate)21/28 = 75.0%
Results
:
see
details by S. Santibanez (RKI)PCR/NAT - Measles Virus Collaboration – WHO EURO and INSTAND (991) Nov 2015
Slide1321.06.1613Evaluation of sequence quality – Measles
Sample 991001: D8-FM- 20 countries/ 21 labs7 countries
The N-450
correctly
sequenced
Deviation(s)
Sample 991004: D4-M
- 23 countries/ 24
labs
4 countriesSample 991003: B3-H- 24 countries/ 25 labs 3 countries- Panel contained three MV-positive samples- Sequence data provided by 27 countries/ 28 labs (for all samples) - All samples correctly sequenced by 20 countries/ 21 labs- Deviations from the reference sequence in:-
one
sample
3 countries - two samples 1 country- three samples 3 countries Incomplete endsMismatch at one or more pos.Single „N“ at one or more pos.Single additional nt insideSingle nt missing inside
Slide14PCR/NAT - Measles Virus Collaboration – WHO EURO and INSTAND (991) Nov 2015 Summary of applied methods and reported ct-values:
ParticipantExtractionAmplification
Sample 991001
Sample 991003
Sample 991004
Sequencing
48512-
IRE
QG Qiamp Viral RNA mini
in house TaqMan
22.9728.7424.58in house48659- UKBiomerieux Easy Magin house Multiplex TM N29.8030.6031.20
no information
48659-
UKBiomerieux Easy Magin house Multiplex TM H30.9035.4032.70no information1296- AUTBiomerieux Easy Magin house real-time PCR26.9026.6025.60Santibanez, JGV 20025577- BULQG Qiamp Viral RNA miniin house PCR (CDC)---not registered f. typing49701- CZEStratecmanufacturer (LifeRiver)28.8428.3529.62not registered f. typing
49690-
DEN
RO MagnaPure 96
in house
TaqMan
39.48
39.41
37.96
in house (WHO)
30010-
DEU
Qiagen
in house
TaqMan
(CDC)
26.27
27.17
26.50
in house
30010-
DEU
Qiagen
in house nested PCR
-
-
-
in house
40790-
EST
no information
in house
TaqMan
(CDC)
21.21
23.15
26.86
in house (CDC)
32637-
FIN
Biomerieux Easy Mag
in house TaqMan
24.76
25.12
24.34
in house
49713-
HUN
Phenol-Chloroform
in house nested PCR
-
-
-
in house (ABI)
49636-
ITA
QG
RNeasy
in house real-time PCR
33.16
27.26
27.77
in house
1191-
LVA
Biomerieux Easy Mag
in house real-time PCR
23.31
22.06
22.78
reminded /no respond
49131-
LTUQG MiniElute Virus Spinmanufacturer (LifeRiver)26.7625.6624.82not registered f. typing40201- NORQG Qiamp Viral RNA miniin house nested PCR---no information49712- POLQG Qiamp Viral RNA miniin house nested PCR---Santibanez, JGV 200249638- ROMMacherey-Nagel Nucleospinin house TaqMan (CDC)24.9626.0225.47in house (ABI)48876- SVNQG MiniElute Virus Spinin house real-time PCR25.7020.8320.74in house (CDC)8850- SWEQG Qiamp Viral RNA miniin house PCR - - -in house (ABI)29411- BELQG Qiamp Viral RNA mini in house TaqMan21.8022.2922.55in house nested PCR49699- BiHQG Qiamp Viral RNA miniin house PCR (CDC)---in house (CDC)49698- CROQiagenin house TaqMan27.0027.0027.00in house49691- FRA CQG EZ1 DSP Virus in house real-time PCR25.9524.9926.63in house49693- GREMacherey-Nagel Nucleospinin house TaqMan26.8825.5824.85In house49709- ISRQiagenin house TaqMan (CDC)26.5022.9022.10in house (CDC)49694- LUXQG Qiamp Viral RNA mini in house real-time nested PCR28.4427.8325.55no information49664- NED BRO MagnaPure 96in house real-time PCR29.6229.8329.83in house49637- NED RRO MP LC total NA isolatioin house real-time PCR22.6022.4021.60in house (CDC)49700- PORBiomerieux Easy Magmanufacturer (FTD)21.5026.4423.63in house nested PCR49692- ESPQiagenin house nested PCR---in house49711- TURQG EZ1 DSP Virusmanufacturer (FTD)21.3030.7021.90in house (BE)49640- BLRQG Qiamp Viral RNA miniin house TaqMan (CDC)25.1324.9925.42in house (CDC)49710- KAZInvitrogen PureLinein house real-time PCR26,6626,2325,62in house (ABI)49710- KAZInvitrogen PureLinein house PCR---in house (ABI)49644- RUSQG Qiamp Viral RNA mini in house real-time PCR25.2023.8023.80in house (ABI)49644- RUSQG Qiamp Viral RNA mini in house PCR---in house (ABI)49697- TJKQG Qiamp Viral RNA mini in house TaqMan26.5026.9027.20not registered f. typing
Red
=
weak
PCR
signals
Blue=
problems
in seq.
quality
Slide15Summary of organizational data:No. of laboratories invited for participation: 37 (call of participation beginning Sep 2015)No. of laboratories registered for participation: 30 (as of 7 Oct 2015)Bulk shipment of sample panels to RRLs: 18.11.2015No. of laboratories reporting results:
30 (from 28 countries)(as of 10 Mar 2016)Response rate: 100%PCR/NAT - Rubella Virus Collaboration – WHO EURO and INSTAND (992) Nov 2015
Slide16Sample properties (as discussed in Aug 2015):PCR/NAT - Rubella Virus Collaboration – WHO EURO and INSTAND (992) Nov 2015 Sample No.
Sample sourceStrain name and sequence ID in RubeNSDilution
992001
Lysate of cells infected with rubella virus
(
wild-type rubella
strain;
genotype
1G
)$RVi/Prahova region.ROU/25.03, 268undiluted992002*= 992004Lysate of cells infected with rubella virus (wild-type rubella strain; genotype 2B)$RVs/Duesseldorf.DEU/35.13, 10074undiluted992003Lysate of non-infected cells----992004*= 992002Lysate of cells infected with rubella virus (wild-type rubella strain;
genotype
2B)$ RVs/Duesseldorf.DEU/35.13, 10074undiluted$ Infectious viruses are chemically inactivated on the sample FTA disk.* The positive samples 992002 and 992004 are identical.
Slide17PCR/NAT - Rubella Virus Collaboration – WHO EURO and INSTAND (992) Nov 2015 Summary of qualitative results and typing results:
& & The success rates for all 4 samples in test categories 20 and 40, respectively, refer to the number of participating laboratories. Laboratories having reported results obtained by several methods are recorded only once.
§ One
participant (Participant No. 1191), who was registered for genotyping, did not report results in test category 40.
This was evaluated as "false" result in test category 40.
# Four
participants (Participant No. 32637, 49638, 49690 and 49711), who were registered for genotyping, were not able to report results in test category 40 due to weak PCR signals. These were evaluated as "false" results in test category 40.
Sample 992001
Sample 992002= 992004Sample 992003Sample 992004= 992002Successratefor all samples&Sample propertiesRubella virus positive(genotype 1G) 104.69 PFU/ml on Vero/hSLAMRubella virus positive(genotype 2B)
10
6.0
PFU/ml on Vero/hSLAMRubella virus negativeRubella virus positive(genotype 2B)106.0 PFU/ml on Vero/hSLAMDilutionundilutedundiluted----undiluted"correct" results for qualitative genome detection (test category 20)(basis for receiving a certificate of successful participation)qual. detectionpositive100% (31/31)positive96.8% (30/31)negative96.8% (30/31)positive100% (31/31)
96.7%
&
(29/30)
&
"
correct
"
results
for
type
identification
(
test
category
40)
(basis for receiving a certificate of successful participation)
type
identification
1G
75.0%
(
18/24)
2B
70.8%
(
17/24)
negative/
nd
75.0%
(18/24)
2B
66.7%
(
16/24)
62.5%
&,§,#
(15/24)
&,§,#
Slide18PCR/NAT - Rubella Virus Collaboration – WHO EURO and INSTAND (992) Nov 2015 & & The success rates for all 4 samples in test
categories 20 and 40, respectively, refer to the number of participating laboratories. Laboratories having reported results obtained by several methods are recorded only once.Link to results from regular INSTAND EQA scheme (389):
Sample 992001
Sample
992002
=
992004
Sample 992003
Sample
992004
= 992002Successratefor all samples&= 389012= 389009= 389011= 389010Sample propertiesRubella virus positive(genotype 1G)Rubella virus positive(genotype 2B)
Rubella virus negative
Rubella virus positive
(genotype 2B)Dilutionundilutedundiluted----undiluted"correct" results for qualitative genome detection (test category 20)(basis for receiving a certificate of successful participation)qual. detectionpositive100% (31/31)positive96.8% (30/31)negative96.8% (30/31)positive100% (31/31)96.7%&(29/30)&qual. detectionpositive94.7%
(18/19)
positive
89.5%
(17/19)
negative
94.7%
(18/19)
positive
89.5%
(17/19)
89.5%
&
(17/19)
&
"
correct
"
results
for
type
identification
(
test
category
40)
(basis for receiving a certificate of successful participation)
type
identification
1G
75.0%
(
18/24)
2B
70.8%
(
17/24)
negative/
nd
75.0%
(18/24)
2B
66.7%
(
16/24)
62.5%
&
(15/24)
&
type
identification
1G
100
%
(1/1)
2B
100
%
(1/1)
negative/
nd
100
%
(1/1)
2B100% (1/1)100%&(1/1)&
Slide19New test categoryfor sequence quality:Success rate for all samples refers
to the number of participating laboratories(in 1st round/Nov 2015 not evaluated w/o consequences for certificate)13/19 = 68.4%
Results
:
see
details by S. Santibanez (RKI)PCR/NAT - Rubella Virus Collaboration – WHO EURO and INSTAND (992) Nov 2015
Slide2021.06.1620Evaluation of sequence quality – Rubella
Sample 992001: 1G- 13 countries/ 14 labs5 countriesSample 992002: 2B- 13 countries/ 14 labs4 countries
1
country
without
result
Sample 992004: 2B
-
13 countries/ 14 labs- 2 countries- 3 countries without resultThe 739 nt window of E1 correctly sequencedDeviation(s)- Panel contained three RV-positive samples- Sequence data provided by 18 countries/ 19 labs: - 15 countries/ 16 labs for all samples - 2 countries for
two
samples - 1 country for one sample - All samples correctly sequenced by 12 countries/ 13 labs- Deviations from the reference sequence in:- one sample 3 countries - two samples 1 country - three samples 2 countriesIncomplete endsMismatch at one or more pos.Gap, stretch of „N“And others:single nt missingsingle add. nt inside- single „N“
Slide21PCR/NAT – Rubella Virus Collaboration – WHO EURO and INSTAND (992) Nov 2015 Summary of applied methods and reported ct-values:
ParticipantExtractionAmplification
Sample
992001
Sample
992002
Sample
992004
Sequencing
48512-
IREQG Qiamp Viral RNA mini in house TaqMan27.4739.8738.40in house48659- UKBiomerieux Easy Magin house TaqMan27.5024.9024.90
no information
1296-
AUTBiomerieux Easy Magin house real-time PCR26.4027.3027.20Fene, SantibanezJMV 20115577- BULQG Qiamp Viral RNA miniin house PCR (CDC)---not registered f. typing49701- CZEStratecmanufacturer (LifeRiver)29.9930.7831.10not registered f. typing49690- DENRO MagnaPure 96in house TaqMan37.7537.7438.07
in house (WHO)
30010-
DEU
Qiagen
in house nested PCR
-
-
-
in house
32637-
FIN
Biomerieux Easy Mag
in house TaqMan
25.51
38.00
36.94
in house
49713-
HUN
Phenol-Chloroform
in house nested PCR
-
-
-
in house (ABI)
49636-
ITA
QG RNeasy
in house real-time PCR
27.76
26.33
27.12
in house
1191-
LVA
Biomerieux Easy Mag
in house real-time PCR
24.69
23.81
25.63
reminded/no respond
49131-
LTU
QG MiniElute Virus Spin
manufacturer (LifeRiver)
27.04
26.12
26.31
not registered f. typing
40201-
NOR
QG Qiamp Viral RNA mini
in house real-time PCR
24.57
23.35
22.51
no information
49712-
POL
QG Qiamp Viral RNA mini
in house nested PCR
-
-
-
Abernatly
,
JCM
2009
49638-
ROM
Macherey-Nagel Nucleospinin house TaqMan (CDC)28.5929.2027.80no info, lab states:PCR signals too weak48876- SVNQG MiniElute Virus Spinin house real-time PCR26.7624.3826.09in house (CDC)29411- BELQG Qiamp Viral RNA mini in house TaqMan25.0226.3226.83not registered f. typing49699- BiHQG Qiamp Viral RNA miniin house TaqMan (CDC)29.2623.9924.03in house (CDC)49696- FRA VBiomerieux Easy Magin house TaqMan26.4027.1026.40in house (ABI)49691- FRA CQG EZ1 DSP Virus in house real-time PCR26.8225.4825.63not registered f. typing49693- GREMacherey-Nagel Nucleospinin house TaqMan26.4225.3925.74In house49709- ISRQiagenin house TaqMan (CDC)26.4024.5024.70in house (CDC)49694- LUXQG Qiamp Viral RNA miniin house TaqMan (modified CDC)29.1025.9425.97no information49664- NED BRO MagnaPure 96in house real-time PCR30.3340.7941.07in house49637- NED RRO MP LC total NA isolationin house real-time PCR21.8032.2031.60in house (CDC)49700- PORBiomerieux Easy Magmanufacturer (Sacace)29.0940.7225.04results disregarded49692- ESPQiagenin house nested PCR---in house49711- TURQG EZ1 DSP Virusin house TaqMan (CDC)32.50neg.-F32.40no information49640- BLRQG Qiamp Viral RNA miniin house TaqMan (CDC)28.6026.4625.73in house (CDC)49644- RUSQG Qiamp Viral RNA mini in house real-time PCR23.3021.2021.87in house (ABI)49644- RUSQG Qiamp Viral RNA mini in house nested PCR---in house (ABI)Red= weak PCR signals Blue= problems in seq. quality
Slide22ConclusionsLink of WHO and INSTAND measles & rubella molecular EQA schemes was successfulSample shipment via WHO EURO and RRLs was smooth (in most cases)Participants were successful in general (success rates 62.5 - 100%)Non-reporters contributed to high failure rateFor future EQA schemes *agreed at 11th Meeting of the Measles/Rubella RRLs WHO EUR – Berlin, 15.03.2016:Inclusion of sequence quality for assessmentEvaluation criteria: 100% correct results in all test categories for all panel members
for labs genotyping: correct genotype and zero nucleotide errorCoordination of sample selection between WHO, CDC and INSTANDHarmonization of next rounds between regional and global EQA schemes (instruction sheet, check-list of required information, evaluation criteria) Online-reporting of EQA results incl. sequence submission to MeaNS/RubeNS (cooperation of PHE with WHO, CDC and INSTAND)Classification of mEQA samples as non-infectious by INSTAND
Slide23Thanks to INSTAND Target Value Laboratories
Robert Koch-Institut, Abt. Infektionskrankheiten, FG 12 Masern, Mumps, Röteln und Viren bei Abwehrschwäche, Nationales Referenzzentrum für Masern, Mumps und Röteln: Prof. Dr. A. Mankertz, Dr. S. SantibanezUniversität Leipzig, Institut für Virologie: Prof. Dr. U. G. LiebertLabor Berlin - Charité Vivantes GmbH, Fachbereich Virologie: Prof. Dr. D. H. Krüger, Prof. Dr. J. HofmannUniversität Würzburg, Institut für Virologie und Immunologie: Dr. B. Weißbrich
Universitätsklinikum Bonn, Institut für Virologie:
Prof. Dr. C. Drosten, Prof. Dr. A.-M. Eis-
Hübinger
Universitätsklinikum Frankfurt, Institut für Medizinische Virologie, Frankfurt/Main, Nationales Referenzzentrum für Retroviren:
Prof. Dr. V.
Kempf
, Prof. Dr. H. F. Rabenau, PD Dr. A. Berger, PD Dr. M. Stürmer
WHO HQ
WHO EUROCDC GSLRRLBerlinPHE GSLLondonRRL LuxembourgRRL MoscowIQVD TeamGBD TeamINSTAND Team
Slide24