Abstract Book of EAVLD2012 - eavld congress 2012

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S3 - P - 03 EVALUATION OF DIRECT BLOOD POLYMERASE CHAIN REACTION FOR RAPID DETECTION OF AFRICAN SWINE FEVER VIRUS Claudia Gabriel 1 , Marianna Khachatryan 1 , Immanuel Leifer 1,2 , Sandra Blome 1 , Johanna Zemke 1 , Bernd Hoffmann 1 , Martin Beer 1 1 Friedrich-Loeffler-Institut, Institute of Diagnostic Virology, Greifswald – Insel Riems, Germany 2 Present address: Institute of Virology and Immunoprophylaxis, Mittelhaeusern, Switzerland Introduction Most polymerase chain reaction (PCR) techniques for the detection of African swine fever virus (ASFV) require labourintensive and expensive DNA extraction steps. Nowadays, PCR kits are available that allow PCR directly from blood samples. Here, we report on the evaluation of such kits in combination with routine primers. Materials & methods In the initial phase of the study, two direct PCR kits were assessed using positive EDTA blood samples obtained from animals experimentally infected with a highly virulent ASFV isolate from Armenia: A) the Phusion® Blood Direct PCR Kit (Finnzymes Oy, Vantaa, Finland; now Thermo Fisher Scientific, Waltham, USA) and B) the KAPA Blood PCR Master Mix (PEQLAB Biotechnologie GmbH, Erlangen, Germany). For specific detection of ASFV, primers published by King et al. (2003) were used. Both test kits were optimised for gel-based PCR regarding sample volume and temperature profile referring to the manufacturer’s recommendations. Subsequently, the direct PCR Kits were tested with negative samples, positive samples from animal trials, and dilution series of positive samples in comparison with the routinely used real-time PCR assay. Results After preliminary optimisation, ASFV was reliably detected in blood samples from experimentally infected pigs. Both with these experimental samples and dilution series of positive samples, the direct PCR proved to be as sensitive as established real-time PCR protocols. Discussion & conclusions Concluding, this assay is favourable in both economical and technological terms and allows the analysis of very small sample volumes. Moreover, direct blood PCR could be advantageous for pen-side diagnostic applications. References 1. King DP, Reid SM, Hutchings GH, Grierson SS, Wilkinson PJ, Dixon LK, Bastos AD, Drew TW. (2003). Development of a TaqMan PCR assay with internal amplification control for the detection of African swine fever virus. J Virol Methods, 107(1):53-61 African swine fever virus, Diagnosis, Direct PCR
S3 - P - 04 DEVELOPMENT OF A SCHMALLENBERG VIRUS ANTIBODY ELISA Christian Schelp 1 , Yann Senechal 1 , San Pun 1 , Daniel Schumacher 1 , Dragos Gradinaru 2 , Christoph Egli 1 , Jean- Luc Troch 1 , John Lawrence 3 , Serge Leterme 3 Introduction Schmallenberg virus was first identified in Germany in late 2011 as a new Orthobunyavirus genetically highly similar to viruses of the Simbu serogroup. The virus infects ruminants such as cattle, sheep and goats. Schmallenberg virus infections have been recently confirmed in several European countries including Germany 1 , The Netherlands, Belgium, France, United Kingdom, Luxembourg, Italy and Spain. Clinical signs include reduced milk yields, fever, diarrhea, abortions and malformed newborns. Virus detection by real time RT-PCR or virus cultivation is not the method of choice for infection monitoring due to the very short viremic period. Therefore, there is an urgent need for an antibody ELISA to identify infected animals. Materials & methods Hundreds of samples originating from clinically affected herds, from virus positive animals and from areas with no Orthobunyavirus presence are being collected. Samples will be further characterized by using an inhouse immunofluorescence assay, PCR and epidemiological and clinical data. An antibody ELISA will be described. Microtiter plates will be coated with inactivated Schmallenberg-virus antigen. Binding of Schmallenbergvirus antibodies will be visualized by colour change in the wells of the microtiter plate. The diagnostic relevance of the result will be assessed by comparing the optical density (OD) of the samples with the OD of the positive control. Results Preliminary data will be presented. Specificity and sensitivity data will be calculated using characterized samples. The data will allow for setting a cut-off to identify Schmallenbergvirus antibody positive and negative samples. Discussion & conclusions The data will be discussed and conclusions will be proposed according to obtained antibody test performance. Acknowledgements We kindly acknowledge the collaboration with the FLI (Friedrich- Loeffler-Institut), Insel Riems, Germany. We also would like to thank for the assistance by various institutes and veterinarians across Europe in helping to collect samples and additional data. References 1. Hoffmann, B, Scheuch, M, Höper, D, Jungblut, R, Holsteg, M, Schirrmeier, H, Eschbaumer, M, Goller, K.V., Wernike, K, Fischer, M, Breithaupt, A, Mettenleiter, T.C., Beer, M (2012). Novel Orthobunyavirus in Cattle, Europe, 2011. Emerging Infectious Diseases, vol. 18, 469-472 1 IDEXX Switzerland AG, Liebefeld, Switzerland 2 IDEXX Montpellier SA, Montpellier, France IDEXX Laboratories, Westbrook, USA 3 Schmallenberg virus, antibody ELISA, Orthobunyavirus
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2 nd CONGRESSOFTHEEUROPEAN ASSOCIAT
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Welcome Dear colleagues, Welcome to
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worked with many diagnostic compani
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Products and services for scientist
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16:4518.15 20:0023:00 [S1.]Oralpres
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Day4 Wednesday,4July2012 4 th sessi
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Oral presentations “General Sessi
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antigens or attenuated vaccines can
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conditions, e.g. Staphylococcus aur
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S1 - O - 1 ORAL FLUIDS - SAMPLE MAT
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S1 - O - 03 INTRODUCTION RATE OF A
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S1 - O -05 VALIDATION OF A COMPETIT
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S1 - O - 07 SUBTYPING OF SWINE INFL
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S1 - O - 09 VIRAL DIAGNOSIS USING T
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S1 - O - 11 USING ORAL FLUID FOR TH
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S1 - O - 13 THE FIRST YEAR OF OBLIG
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S1 - O - 15 SEROLOGICAL ANALYSIS AN
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S1 - O - 17 RING TEST EVALUATION FO
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Oral presentations “Diagnostics a
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S2 - O - 01 DEVELOPMENT OF A RAPID
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S2 - O - 03 EVALUATION OF RAPID HRS
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Oral presentations “Emerging, re-
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advances, the fundamental tools of
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S3 - O - 02 25 YEARS OF PASSIVE SUR
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S3 - O - 04 EQUINE NOCARDIOFORM PLA
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S3 - O - 06 DETECTION OF SCHMALLENB
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S3 - O - 08 SCHMALLENBERGVIRUS: SER
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S3 - O - 10 DIAGNOSTIC ASPECTS OF S
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Oral presentations “New technique
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acid and organic solvent. Then a dr
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S4 - O - 02 BIOLOG GENERATION III,
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Page 67 and 68: S4 - O - 06 15 MINUTE ELISA USING A
Page 69 and 70: S1 - P - 01 LAWSONIA INTRACELLULARI
Page 71 and 72: S1 - P - 03 DETECTION OF PRRSV ANTI
Page 73 and 74: S1 - P - 05 DEVELOPMENT AND EVALUAT
Page 75 and 76: S1 - P - 07 ANALYTICAL EVALUATION O
Page 77 and 78: S1 - P - 09 PTS FOR BVDV ANTIGEN DE
Page 79 and 80: S1 - P - 11 ANTIMICROBIAL SUSCEPTIB
Page 81 and 82: S1 - P - 13 IDENTIFICATION OF GLOBI
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Page 85 and 86: S1 - P - 17 VIROLOGICAL SURVEILLANC
Page 87 and 88: S1 - P - 19 CHARACTERIZATION OF EXT
Page 89 and 90: S1- P - 21 ANALYSIS OF THE EFFICIEN
Page 91 and 92: S1 - P - 23 RELIABLE AND EARLY DETE
Page 93 and 94: S1 - P - 25 GENOTYPIC VARIABILITY D
Page 95 and 96: S1 - P - 27 PROFICIENCY TESTING (PT
Page 97 and 98: S1 - P - 29 LOW PATHOGENIC AVIAN IN
Page 99 and 100: S1 - P - 31 PRRSV DIAGNOSTICS BY RT
Page 101 and 102: S1 - P - 33 APPLICATION OF A HERD P
Page 103 and 104: S1 - P - 35 DEVELOPMENT OF A BROADL
Page 105 and 106: S1 - P - 37 MONITORING OF BOVINE VI
Page 107 and 108: S1 - P - 39 BOVINE BLOOD DENDRITIC
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Page 119 and 120: S3 - P - 06 COMPARISON OF CULTURE A
Page 121 and 122: S3 - P - 08 EMERGENCE OF SCHMALLENB
Page 123 and 124: S3 - P - 10 SIMULTANEOUS DETECTION
Page 125 and 126: S3 - P - 12 PRELIMINARY VALIDATION
Page 127 and 128: S3 - P - 14 DEVELOPMENT OF MULTISPE
Page 129 and 130: S3 - P - 16 DETECTION OF IBV QX IN
Page 131 and 132: S3 - P - 18 PREVALENCE OF COXIELLA
Page 133 and 134: S3 - P - 20 INTERFERON-GAMMA ASSAY
Page 135 and 136: S3 - P - 22 COMPARISON OF THE PERFO
Page 137 and 138: S3 - P - 24 DEVELOPMENT OF A VIRUS
Page 139 and 140: S3 - P - 26 DETECTION OF EQUINE FOA
Page 141 and 142: S4 - P - 01 MOLECULAR CHARACTERIZAT
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Page 151 and 152: S4 - P - 11 EFFECTIVE TESTING STRAT
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Page 155 and 156: S1-P-03 BALLAGI ANDREA S1-P-04 BENI
Page 157 and 158: S3-P-06 KELLER SELINA S3-P-07 KÖNI
Page 159 and 160: List of abstracts (alphabetical ord
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Abstract Book of EAVLD2012 - eavld congress 2012

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