Tasks of the European Network for diagnostics of 'Imported ...

Tasks of the European Network for diagnostics of 'Imported ...

West Nile Source: H. Gelderblom, RKI First international proficiency study on West Nile virus molecular detection. ROBERT KOCH INSTITUT Matthias Niedrig, Sonja Linke Christian Drosten Herv Zeller ENIVD 06/06 10:58 10:58 Distribution of disease vectors and WN virus in Europe Aedes albopictus Ochlerotatus atropalpus WN Ochlerotatus japonicus WN WN source: Snow & Ramsdale, Biologist (2002) 49 (2) Hubalek & Halauzka; EID, (1999) 5 (5) ENIVD 05/06 10:58 10:58 WN WN WN WN Import of suspected or confirmed WN cases into Europe eight cases in total Viral agent Number of cases/deat hs Type of traveller Reference Switzerland Ebola virus 1/0 Repatriation 7 Brazil Switzerland Yellow Fever virus 1/1

Tourist 8 Sep 1997 Ghana Germany Suspected VHF 1 1/1 Refugee 9 Nov 1998 The Gambia Belgium Suspected VHF 2 1/1 Tourist 10 Aug 1999 Ivory Coast Germany Yellow Fever virus 1/1 Business 11 Jan 2000 i.a. Ivory Coast Germany Lassa virus 1/1 Tourist 12 Feb 2000 Sierra Leone UK Lassa virus 1/1 Repatriation 13 Mar 2000 Nigeria Germany

Lassa virus 1/1 Medical transport 14 Jul 2000 Sierra Leone The Netherlands Lassa virus 1/1 Business 15 Dec 2000 Kenya Germany Suspected VHF3 1/1 Tourist 16 Mar 2001 Sierra Leone Germany Suspected VHF2 1/0 Repatriation 17 Mar 2001 i.a. Chile France Hantavirus 1/0 Tourist 18 Aug 2001 Bulgaria Germany CCHF virus 1/0 Tourist Pers. comm. a) Sep 2001 Chad France

RVF virus 2/0 Business 19 Nov 2001 The Gambia Belgium Yellow Fever virus 1/1 Tourist 20 Sep 2002 Nepal Spain Suspected VHF 1 1/0 Tourist Pers. comm. b) Oct 2002 Cameroon Rep.of Ireland Suspected VHF 2 1/0 Business Pers. comm. c) Feb 2003 Sierra Leone UK Lassa virus 1/0 Business 21 Feb 2003 i.a China different countries SARS-CoV 33/1 Tourist/Business 6 Mar 2003

USA France West Nile virus 1/0 Tourist Pers. comm. d) Aug 2003 USA Germany West Nile virus 2/0 Tourist 22 Jun 2004 USA France Hantavirus 1/1 Tourist Pers. comm. d) Sep 2004 USA Germany West Nile virus 1/0 Tourist 22 Oct 2004 USA France West Nile virus 3/0 Tourist Pers. comm. d) Oct 2004 Tunisia France West Nile virus 1/0 Tourist Pers. comm. d) Nov 2004

Senegal CCHF virus 1/0 Repatriation Pers. comm. d) Period of time Imported from Imported to Nov 1994 Ivory Coast Apr 1996 ENIVD 05/06 France 10:58 10:58 Preparation of PCR samples for the External Quality Assurance (EQA) cell culture supernatants of infected cells with the respective viruses heat treatment for 1h at 56C and gamma irradiation of 30 kGy check for infectivity by cell cultivation dilute with human plasma, aliquots of 100 l are frozen at 70C before freeze-drying for 12 hours samples were distributed by normal mail service ENIVD 05/06 10:58 10:58 Phylogenetic tree of some tested WNV samples and reference strains Source: Alex Platonov 0.01 based on the UPGMA method by using WN-HNY1999/AF202541/ WN-IS-98-STD-STORK/AF481864/ WN-NY99-F382-99/AF196835/ WN-MNY-2741/AF206518/ WN-NY99-eqhs/AF260967/ 12 ENIVD 9 ENIVD 2 ENIVD AY278441 Ast99-901 WN-NY2000-grouse3282/AF404755/ 4 ENIVD WN-Eg101/AF260968/ WN-ITALY98-EQUINE/AF404757/ AY490240 Chin strain WN-Volgograd-1999 AF317203 WN-Romania-1996M/AF260969/ AY262283 KN3829 C univittatus 5 ENIVD AY268132 PaAn001 AY268133 PaH001 Kunjin-MRM61C/D00246/ AY274505 Kunjin Pakun WN-Sarafend/AY688948/ WN-B956/AY532665/ 1 ENIVD WN-WNFCG/M12294/

6 ENIVD AY277251 LEIV-Krnd88-190 D marginatus ENIVD 05/06 10:58 nt 10:58 216 sequence of 5'-UTR/C gene fragment Evaluation of the WN-PCR EQA laboratory n origin copy N [ge/ml] samples n #5 #1 WN France WN Uganda #2 WN-NY #9 WN-NY #12 WN-NY #4 WN-NY #6 Kunjin 9,4E+05 9,4E+04 9,4E+03 9,4E+02 1,0E+06 1,0E+03 1,0E+03 3 5 2 3 6 5 3 8,7x10 2,0x10 2,7x10 7,8x10 2,0x10 4,3x10 2,6x10 1 7 5 5 4 6 6 1,2x10 7,8x10 1,5x10

3,6x10 8x10 1,37x10 2 5 WN 1 WN 1 WN 1 WN-1 WN 1 WN 2 WN 6 WN-NY WN-NY WN-NY WN-NY WN WN WN 7 + + + + + (+) + 19 + + + + + + 4 + + + + + WN 2 WN-2 6 4 3 3 6 5 2 WN ~10 27* WN 1,2x10 WN 3,4x10 WN 3,4x10 WN 3,4x10 WN 1,4x10 WN 1,7x10 (-) 13 + + + + + + (-) 26 WN WN WN WN WN WN 6 5 3 3 4 (-) 1,5x10 1,5x10 1,5x10 1,5x10 1,5x10 21 6 5 4 3

6 (-) 3,8x10 5,6x10 7,4x10 5,1x10 7,7x10 18 (-) 22 + + + + + (-) 23 + + + + + (-) 15 + + + + + (-) 3 + + + + + (-) 31 + + + + + (-) 32 WN-NY WN-NY + WN-France WN-Uganda + (-) 14 + + + + + (-) 33 + + + + + + (-) (-) 20 + + + + 4 3 2 2 5 (-) (-) 1x10 WN-2 1,7x10 16 WN1 4,5x10 WN1 8,5x10 WN1 4,2x10 (-) (-)

(-) 8 + + + (-) (-) (-) 10 + + + (-) (-) (-) 30 + + + (-) (-) (-) 29 + + + (-) (-) (-) 37 + + + ENIVD 05/06 (-) (-) (-) (-) 24 + + 10:58 10:58 (-) (-) (-) (-) (-) (-) 34 - #10 #11 #3 #7 #8 SLE / JEV YF-17D/TBE Dengue 1-4 neg. neg. 1,0E+03 1,0E+03 1,0E+03 - SLE - SLE WN (+) WN (+) - - YF - Flavi 2 WN ~10 (+) - - DEN 3 - DEN 2 - DEN 2 - DEN - DEN 2 n.d. - DEN WN (+)

- - +/- correct Score results in % 11 11 11 11 11 11 11 10 10 10 10 10 10 10 10 10 10 10 9 9 9 8 8 8 8 8 7 7 5 100,0 100,0 100,0 100,0 100,0 100,0 100,0 90,9 90,9 90,9 90,9 90,9 90,9 90,9 90,9 90,9 90,9 90,9 81,8 81,8 81,8 72,7 72,7 72,7 72,7 72,7 63,6 63,6 45,5 Evaluation of WN PCR diagnostic by EQA (1) WNV-1 Nof laboratories % copy N [ge / ml] 100.000 28 96,6 10.000

26 89,7 1.000 22 75,9 100 19 65,5 10 34,5 4 2,9 WNV-2 YF, SLE, DEN 1 - 4, TBE, neg. ENIVD 05/06 10:58 10:58 Evaluation of the WNV-PCR diagnostic by EQA (2) 1,0E+08 control Reihe1 lab. 1 Reihe2 lab. 2 Reihe3 lab. 21 Reihe5 lab. 16 Reihe6 lab. 18 Reihe7 copy N [ge/ml] 1,0E+07 1,0E+06 1,0E+05 1,0E+04 1,0E+03 1,0E+02 1 #2 2 #9 sam ple ENIVD 05/06N 10:58 10:58 3 # 12 4 #4 lab.

27 Reihe4 Fraction of laboratories Evaluation of WNV PCR diagnostic by EQA (3) lineage 1 lineage 2 Log RNA copies / ml ENIVD 05/06 10:58 10:58 Probit analysis of the fractions of laboratories achieving positive results (y-axis) in relation to the virus RNA concentration in a given positive sample (xaxis). Data points represent individual samples in the proficiency test panel. The thick line is the regression line calculated on the basis of a probit model (doseresponse curve), the thin lines are 95% confidence Results WNV PCR EQA Does the PCR method used has an influence on the quality of the results? n= 28; 15 real time PCR, 11 RT-PCR, 2 commercial There is no significant correlation between the PCR method and the quality of the results. Do other factors influence the PCR results ? genome preparation: n= 28, 19 Qiagen, 5 Roche, 4 other There is no significant correlation between the genome preparation method and the quality of the results. ENIVD 05/06 10:58 10:58 - Conclusions - WNV PCR EQA using cell culture supernatants, diluting samples with human plasma and freeze-drying before shipping is useful for sample preparation - gamma-irradiation is a suitable method for inactivation of samples 7 laboratories showed excellent results (sensitivity of 100%), whereas 10 laboratories that were less sensitive need to improve their methods PCR methods for detection of WNV should also detect lineage 2 which is now present in Europe. 4 laboratories create false positive results ENIVD 05/06 10:58 10:58 EQA studies performed by the ENIVD between 1999 and 2005 Period of time Viral agent 1999/2000* 2001/2002 1999* 2002 2002/2003 2002/2003 2002/2003 2002/2003 2004 2004 2004/2005 2005 2005 2005 2005

2006 Hantavirus Hantavirus Dengue virus Dengue virus Dengue virus Filovirus Lassa virus Orthopox virus SARS-CoV SARS-CoV Orthopox virus West Nile Virus West Nile Virus Tick borne Enc. Tick borne Enc. Yellow Fever No. of No. of No. of labs with Type of No. of samples participants participants good overall methods positive/negative (labs) (countries) proficiency Serology 13 10 3/1 11 (85 % ) Serology 18 14 14/6 13 (72 % ) Serology 13 10 2/2 11 (85 % ) Serology 18 16 18/2 15 (83 % ) PCR 13 12 7/3 5 (38 % ) PCR 14 13 7/5** 7 (50 % ) PCR 14 13 8/5** 8 (57 % ) PCR 23 15 13/5** 10 (43 % ) PCR 62 37 7/3 54 (87 % ) Serology 30 19 5/6 13 (43 % ) PCR 34 18

11/9 85% / 58% *** PCR 28 20 6/5 17 (60% ) Serology 28 20 4/6 20 (71% ) PCR 23 16 9/3 9 (39% ) Serology 42 25 8/5 25 (60% ) PCR / Serology study planed Reference published published published published published published published published published published published submitted submitted submitted submitted * Pre-evaluation panel tested before running the respective EQA to optimise sample preparation and shipping procedures. ** The same negative samples were included in the three test panels for diagnostic of Filo-, Lassa- or ENIVD 05/06 10:58 10:58 and specificity including inhibiting factors Orthopox virus. *** consist out of two panels: one for sensitivity Whats coming next ENIVD 05/06 10:58 10:58 Acknowledgement: H. Bin, Central Virol. Laboratory, Sheba Medical Center, Tel Hashomer, Israel D. J. Gubler, CDC, Fort Collins, USA A. Platonov, Central Research Institute of Epidemiology, Moscow, Russia H. Zeller, Jean Mrieux-INSERM P4 laboratory, Institut Pasteur, Lyon, France C. Drosten, Bernhard Nocht Institut, Hamburg, Germany Stephan W. Aberle, Franz Heinz, Medical University of Vienna, Austria Parts of this project were supported by the European Network for diagnostics of "Imported" viral Diseases (ENIVD) presently funded by DG-SANCO of the European Community under the program AIDS and other communicable diseases ENIVD 05/06 10:58 10:58 European Network for diagnostics of "Imported" viral Diseases (ENIVD) for further information: www.enivd.de www.enivd.net www.enivd.org ENIVD 06/06 10:58 10:58

Recently Viewed Presentations

  • Adaptive Witness Encryption and Asymmetric Password-based Cryptography Mihir

    Adaptive Witness Encryption and Asymmetric Password-based Cryptography Mihir

    : IND-CPA adversary A can choose its m. 0 and m 1, but WE adversary B doesn't have that choice Reduction doesn't workFix: Consider a non-uniform B. B can run A with the best coins, which are given to B...
  • 7 Day Skin Care Challenge Business Flow What

    7 Day Skin Care Challenge Business Flow What

    Herbalife SKIN is our new skin care range and it's been clinically tested to show it gets results in 7 Days*! 7 day's use of selected products and skin looks younger and healthier*. You can invite your customers to try...
  • www.adultfinancialed.org

    www.adultfinancialed.org

    Session One. February 25, 2016. Scottsdale Community College. Introduction. We want to sincerely thank you for your time and effort to participate in this workshop. Our hope is th
  • Activity 2.1.1 Creating a Prototype

    Activity 2.1.1 Creating a Prototype

    Selective Laser Sintering (SLS) A rapid prototyping process that used a moving laser beam to sinter heat-fusible powders one layer at a time. General process: A CO 2. laser beam traces shapes of individual slices across a layer of heat-fusible...
  • Force Directed Placement: GPU Implementation Bernice Chan Ivan

    Force Directed Placement: GPU Implementation Bernice Chan Ivan

    Force Directed Placement: GPU Implementation Bernice Chan Ivan So Mark Teper Force Directed Placement Nodes represented by masses Edges represented by springs Motions of elements are simulated iteratively until steady state is reached Basic Implementation The Force Directed Placement algorithm...
  • An Algorithm of Eye-Based Ray Tracing on MATLAB

    An Algorithm of Eye-Based Ray Tracing on MATLAB

    Ray tracing is a special rendering algorithm in 3D computer graphics. 1. Introduction. 1. 2 Ray Tracing Method. ... This model is an additive color model that adds red, green and blue light together to produce other colors. 3. Simple...
  • GEOCENTRIC vs. HELIOCENTRIC - Wine Production

    GEOCENTRIC vs. HELIOCENTRIC - Wine Production

    Our Solar System consists of a central star (the Sun), the nine planets orbiting the sun, moons, asteroids, comets, meteors, interplanetary gas, dust, and all the "space" in between them. The nine planets of the Solar System are named for...
  • High Pressure Vessels - cwilliams.chem.ox.ac.uk

    High Pressure Vessels - cwilliams.chem.ox.ac.uk

    High Pressure Vessels. No lone working is permitted . Be sure that the CO. 2. cylinder is securely fastened.. Periodical leak test in the CO. 2. regulator, lines and reactor.. Use personal protective equipment PPE (specs, gloves, coat).