Front Page Partners Objectives Methodology Project Workplan Background of the proposal Expected benefits and Achievements Contribution Innovation aspects Results News Guestbook
 
 
· Front page
· Partners
· Objectives
· Methodology
· Project Workplan
· Background of the proposal
· Expected benefits and achievements
· Contribution
· Innovation Aspects
· Results
· Exploitation and dissemination activities
· Conclusions
· News
· Duration of the project
· Contact
· Acknowledgements
· Work after the project was closed

Web site. The consortium constructed and launched a web site on the Internet that is regularly updated with results and information (www.multiplex-eu.org). The site has had a total number of 12 025 visits until 30 April 2004, i.e., the closing time of the Final Report. An increasing tendency is seen, since 4 735 visits were recorded under 2003 and 1450 visits in the first four months of 2004. The recorded visits have arrived from 95 countries, a high number of domains, like educational, commercial, government, US government, USA Military, by using a wide range of search engines worldwide. The site has an open web-forum, where the co-ordinator is answering the questions of the visitors and is regularly disseminating the required information. The web-site is kept open after termination the project and will be regularly updated with further results, publications and other news.

Workshop. After publishing in periodical journals the selected methods (molecular beacon, PriProET and Invader) will be made accessible for all interested diagnostic laboratories. From this respect the initial try to transfer technology have already been done inside the Consortium. A hands-on-workshop of PriProET method was organised by Partner 3 (Lindholm, Denmark) with participants from Partner 1, Partner 2, Partner 4 and Partner 5. The main aim of the workshop was to adapt PriProET system to the other types of real-time PCR instruments. Obtained results showed that the system is flexible and can be successfully used on different types of real-time PCR instruments.

 

OIE Reference Laboratory for the Application of Polymerase Chain Reaction Methods for Diagnosis of Viral Diseases in Veterinary Medicine

Considering the importance of the international standardisation of the PCR diagnostics, the coordinator institute (Partner 1) submitted an application for the Office International des Epizooties (OIE) Reference Laboratory status in January 2004. The OIE Biological Standards Commission reviewed the request to designate the Swedish National Veterinary Institute (SVA), Uppsala, as an “OIE Reference Laboratory for the Application of Polymerase Chain Reaction Methods for Diagnosis of Viral Diseases in Veterinary Medicine”. The Commission agreed that SVA has a great deal of expertise in this area, and therefore accepted the application. The International Committee of the OIE approved the supported application during the General Session in May 2004. The honouring nomination of the co-ordinator institute, as the first OIE reference laboratory in molecular diagnostics, is certainly an important outcome of this project.

 

Publications and congress presentations emanating from the project:
(The list is given in alphabetic order)

Agüero, M.; Fernández, J.; Romero, L.; Zamora M.J.; Sánchez, C.; Belák, S.; Arias, M.; Sánchez-Vizcaíno, J.M. 2004. A highly sensitive and specific gel-based multiplex RT-PCR assay for the simultaneous and differential diagnosis of African Swine Fever and Classical Swine Fever in clinical samples. Veterinary Research, 35 (5): 551-563.

Agüero, M; Fernández, J; Arias, M; Romero, L; Canals, A; Sánchez-Vizcaíno, J.M. 2001. “African Swine Fever Virus detection by PCR”. Annual Meeting of National Swine Fever Laboratories. 17-18 May, 2001, Lindholm, Denmark.

Agüero, M; Fernández, J; Romero, L; Sanchez, C; Arias, M; Sánchez-Vizcaíno, J.M. 2003. “Highly sensitive PCR assay for routine diagnosis of African Swine Fever Virus in clinical samples”, J. Clin. Microbiol., 41 (9): 4431-4434.

Agüero, M; Fernández, J; Romero, L; Zamora, M.J; Sanchez, C; Belák, S; Arias, M; Sánchez-Vizcaíno, J.M. 2004. “A highly sensitive and specific gel-based multiplex RT-PCR assay for the simultaneous and differential diagnosis of African Swine Fever and Classical Swine Fever”, Vet. Res., accepted March 18th 2004.

Belák, S. 2003. Molecular epizootiology of positive strand RNA viruses. First CISA Symposium on Animal Health, Valdeolmos, Madrid, Spain, April 10-12, 2003.

Belák, S. 2004. QLK2-CT-2000-00486: Development, standardisation and harmonisation of novel multiplex nucleic acid tests for the detection of economically important viruses of farm animals. Workshop: EU funded research in foot and mouth disease and classical swine fever. EC Commission, Brussels, 22-23 January 2004.

Belák, S. 2004. The molecular diagnosis of porcine viral diseases. Acta Veterinaria Hungarica, 2005. 53. 113-124.

Belák, S., Thorén, P. 2001. Molecular diagnosis of animal diseases. Expert Review of Molecular Diagnostics, 1: 434-444.

Belák, S., Thorén, P. Standardisation of molecular diagnosis of animal diseases. Tenth International Symposium of Veterinary Laboratory Diagnosticians and OIE Seminar on Biotechnology, Salsomaggiore, Parma, Italy, July 4-7, 2001, 3.

Belák, S., Hakhverdyan, M, Thorén, P. Advances in diagnostic techniques. 4th International Symposium on Emerging and Re-emerging Pig Diseases, Rome, Italy, June 29 - July 2, 2003, 19.

Belák, S., Hakhverdyan, M, Thorén, P. Molecular diagnosis of animal diseases: Recent developments and view. 5th National Congress of S.I.Di.L.V., Pisa, 20-21 November 2003. Key lecture.

Fernández, J; Agüero, M; Arias, M; Romero, L; Sánchez-Vizcaíno, J.M. 2002. “Multiplex RT-PCR assay for differential detection of Classical swine fever virus (CSFV) and African swine fever virus (ASFV)”. 5th ESVV Pestivirus Symposium, 26-29 August 2002, Cambridge, United Kingdom.

Fernández, J; Agüero, M; Romero, L; Vazquez, B; Belák, S; Arias, M; Sánchez-Vizcaíno, J.M. 2003. “A highly sensitive and specific one-step multiplex RT-PCR assay for simultaneous and differential diagnosis of animal vesicular diseases”. 6th International Congress of Veterinary Virology, 24-27 August 2003, Saint-Malo, France.

Fernández, J; Agüero, M; Romero, L; Vazquez, B; Belák, S; Arias, M; Sánchez-Vizcaíno, J.M. 2003. "Rapid and differential diagnosis of Foot-and-mouth disease virus, Swine vesicular disease virus, and Vesicular stomatitis virus by one-step multiplex RT-PCR assay in clinical samples". Session of the Research Group of the Standing Technical Committee of the European Commission for the Control of Foot-and-Mouth disease, 16-19 September, 2003, Gerzensee, Berne, Switzerland.

Germán P., Ursu K., Arias M., Kiss I., Belák S., Uttenthal Å., Rasmussen T.B. Adaptation of the ASFV detecting PriProET PCR assay to a BIORAD iCycler instrument. Annual meeting of Hungarian Society for Microbiology, Oct. 7-9. 2004. Keszthely, Hungary. Oral presentation.

Germán P., Ursu K., Arias M., Kiss I., Belák S., Uttenthal A., Rasmussen T.B. 2004.Adaptation of an ASFV detecting PriProET PCR assay to a BIORAD iCycler instrument. Congress of the Hungarian Society for Microbiology, 7-9 October 2004, Keszthely, Hungary.

Hakhverdyan M., Thorén P., Belák S. 2002. Real-time PCR assays for the detection of swine vesicular disease virus and vesicular stomatitis virus using molecular beacons. IUMS World Congress “The World of Microbes”, Abstracts of the XII International Congress of Virology, Paris, 27th July to 1st August 2002, p. 228.

Hakhverdyan M. 2002. Development, standardisation and harmonisation of novel multiplex nucleic acid tests for the detection of economically important viruses of farm animals. Classical Swine Fever Virus Annual Meeting, 2002, Weybridge, UK. Oral presentation.

Hjertner B., Meehan B., McKillen J., McNeilly F. and Belák S. Development of an invader squared assay for the detection of African swine fever DNA. Manuscript.

Jovita Fernández. "Development and standardisation of multiplex PCR assays for the differential diagnosis of economically important infectious animal diseases". PhD thesis. Universidad Autónoma de Madrid, Spain, December, 2005.

Rasmussen, T.B. 2005.Molecular detection of exotic viral infections in animals using quantitative real-time RT-PCR. PhD thesis, Royal Veterinary and Agricultural University, October 2005, Copenhagen, Denmark.

Rasmussen T.B., Uttenthal Å. 2004. Quantitative FRET-based PCR detection of exotic viruses in a multiplex setup: PriProET. Experiences from an EU project. Oral presentation at Stratagene QPCR seminar, Copenhagen, June 2004.

Rasmussen T. B., Uttenthal Å., Fernández J., Storgaard T. Quantitative multiplex assay for simultaneous detection and identification of Indiana and New Jersey serotypes of Vesicular Stomatitis Virus. Journal of Clinical Microbiology. Submitted in April 2004.

Rasmussen,T.B., Uttenthal,Å., Fernandez,J., and Storgaard,T. 2005. Quantitative multiplex assay for simultaneous detection and identification of Indiana and New Jersey serotypes of vesicular stomatitis virus. J. Clin. Microbiol. 43, 356-362.

Rasmussen T.B., Uttenthal Å., de Stricker K., Belák S. and Storgaard T. 2003. Development of a novel quantitative real-time RT-PCR assay for the simultaneous detection of all serotypes of Foot-and-mouth disease virus. Arch. Virol. 148: 2005-2021.

Rasmussen T.B., Uttenthal Å., de Stricker K., Belák S. and Storgaard T. 2004. Detection of foot-and-mouth disease virus in clinical samples using a novel real-time RT-PCR assay. Poster at the European Society for Clinical Virology (ESCV), 15-17 January 2004, Copenhagen, Denmark.

Sámi L., Kiss I., Ursu K., McKillen J. and Prof. S. Belák. Simultaneous detection of PPV, PRRSV and ADV in a multiplexPCR system. Manuscript.

Sámi L., Kiss I., Ursu K., McKillen J. and Prof. S. Belák. 2004. Simultaneous detection of PPV, PRRSV and ADV in a multiplex PCR system. Annual meeting of Hungarian Society for Microbiology, Oct. 7-9. 2004. Keszthely, Hungary. Poster presentation.

Uttenthal Å. 2003. Detection of African Swine Fever virus by quantitative PCR methods. Oral presentation at the national African swine fever meeting in Brussels, May 2003.

Uttenthal, Å., Jensen T.M., Rasmussen, T.B. 2005.Uniform extraction of RNA and DNA for quantitative multiplex PCR detection of classical and African swine fevers in blood or meat juice. Abstract ESVV Pestivirus Symposium, Thun, Schweiz.

Uttenthal Å., Rasmussen T. B. 2004. Real-time multiplex PCR for the detection of African and classical swine fever virus. Oral presentation at the Annual ASF meeting, Grange, Dublin, May 2004.


Conference – oral presentations:

Hakhverdyan M., Rasmussen, T.B., Uttenthal, Å., Thóren, P., Bélak,S. 2005. Experiences on the application of various nucleic acid amplification systems for the improved diagnosis of vesicular diseases”. New Diagnostic Technology: Applications in Animal Health & Biologics Controls. Applications in disease surveillance, molecular epidemiology and quality control tests of vaccines. 3-5 October 2005, Saint-Malo, France. Keynote lecture.

German P. 2005. Detection of Avian influenza virus and Newcastle disease virus by a novel real-time RT-PCR using light upon extension (LUX) fluorogenic primers”. Annual Meeting of the Hungarian Society for Microbiology, jointly organized with the 1st Central European Forum for Microbiology (CEFORM). October 26-28, 2005, Keszthely, Hungary. Oral presentation.

Uttenthal, Å., Rasmussen, T.B., Thóren, P., Bélak,S. 2005. The standardisation, quality control and validation of molecular diagnostic tests. 7th OIE/WAVLD Seminar on Application of biotechnology to Zoonotic Disease Diagnosis. Montevideo, Uruguay. Keynote lecture.

Uttenthal Å., Rasmussen T. B. 2004. Real-time multiplex PCR for the detection of African and classical swine fever virus. Annual ASF meeting, Grange, Dublin, May 2004. Oral presentation.

Posters:

Hakhverdyan M, Rasmussen TB, Uttenthal Å, Belák S. 2005. Detection of swine vesicular disease virus (SVDV) using PriProET Real-Time PCR assay. IUMS, XIII International Congress of Virology, San Francisco, CA, USA, July 23-28, 2005:140-141. Partners involved: P1 (SVA), P3 (DFVF).

Hjertner B.,Meehan B., McKillen J., McNeilly F., Belák S. 2005. Development and evaluation of an INVADER assay for the detection of African swine fever DNA. New Diagnostic Technology: Applications in Animal Health & Biologics Controls. Applications in disease surveillance, molecular epidemiology and quality control tests of vaccines. 3-5 October 2005, Saint-Malo – France. P. 80. Partners involved: P2 (QUB), P1 (SVA).

McKillen J., Brawley A., Hjertner B., McNeilly F., Allan G. 2005. Development of molecular beacon assay for rapid real-time polymerase chain reaction detection of four economically important viruses of swine. New Diagnostic Technology: Applications in Animal Health & Biologics Controls. Applications in disease surveillance, molecular epidemiology and quality control tests of vaccines. 3-5 October 2005, Saint-Malo – France. P. 86. Partner involved: P2 (QUB).