-Equine herpesvirus types 2 and 5 (EHV-2 and EHV-5) have a rather unclear pathogenicity and distribution within the equid population. In order to gain more information on the prevalence of these two viruses, type-specific PCR assays were developed to detect viral DNA in nasal specimens and in peripheral blood leukocytes (PBLs) of adult horses and foals from various regions of Europe, i.e. Sweden, Hungary and the United Kingdom. In adult horses, the prevalence of EHV-2 in PBLs was up to 68% in Sweden and 71% in the United Kingdom. EHV-2 DNA was detected in the PBLs from all the foals tested in all countries and most (93%) of the nasal specimens also yielded positive results. The prevalence of EHV-5 DNA in the PBLs of foals in Hungary was 15 and 24% in adult horses in the United Kingdom. This observation was among the very few reports of the presence of EHV-5 in horses. In summary, the specific PCR assays revealed important data on the occurrence and distribution of EHV-2 and EHV-5 in large horse populations. The findings indicated that infection with EHV-5 occurred later than EHV-2 in foals. This study may contribute to a better understanding of the etiological role of these gammaherpesviruses in equine diseases. Chez les chevaux adultes, la prévalence de EHV-2 dans les LSP atteignait 68 % en Suède et 71 % au Royaume-Uni. L'ADN de EHV-2 a été détecté dans les LSP de tous les poulains testés, et la plupart (93 %) des prélèvements nasaux étaient également positifs. La préva-lence de l'ADN de EHV-5 dans les LSP des poulains en Hongrie était de 15 % et de 24 % chez les chevaux adultes au Royaume-Uni. Cette observation fait partie des très rares signalements de la présence de EHV-5 chez les chevaux. En résumé, les tests PCR spécifiques ont révélé des données importantes sur la présence et la distribution de EHV-2 et EHV-5 dans d'importantes populations de chevaux. Les résultats ont montré que l'infection par le EHV-5 se produisait plus tard que celle par le EHV-2 chez le poulain. Cette étude apporte une meilleure compréhension du rôle étiologique de ces herpesvirus gamma dans les maladies équines.infections équines / Gammaherpesvirinae / EHV-2 / EHV-5 / PCR

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Background: Nucleic acid amplification allows the detection of single infectious agents. Protein-based assays, although they provide information on ongoing infections, have substantially less detection sensitivity. Methods: We used proximity ligation reactions to detect proteins on bacteria and virus particles via nucleic acid amplification. Antibodies recognizing viral or bacterial surface proteins were equipped with DNA strands that could be joined by ligation when several antibodies were bound in proximity to surface proteins of individual infectious agents. Results: Detection sensitivities similar to those of nucleic acid-based detection reactions were achieved directly in infected samples for a parvovirus and an intracellular bacterium. Conclusions: This method enables detection of ligated DNA strands with good sensitivity by real-time PCR and could be of value for early diagnosis of infectious disease and in biodefense.

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King, et al.. Evaluation of a novel proximity ligation assay for the sensitive and rapid detection of foot-and-mouth disease virus. Veterinary Microbiology, Elsevier, 2008, 127 (3-4) This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. A novel proximity ligation assay (PLA) using a pan-serotype reactive monoclonal antibody 21 was developed and evaluated for the detection of foot-and-mouth disease virus (FMDV) in 22 clinical samples collected from field cases of disease. The FMDV-specific PLA was found to 23 be 100 times more sensitive for virus detection than the commonly used antigen capture-24 ELISA (AgELISA). As few as 5 TCID 50 were detected in individual assays, which was 25

BackgroundLawsonia intracellularis is a common cause of chronic diarrhoea and poor performance in young growing pigs. Diagnosis of this obligate intracellular bacterium is based on the demonstration of the microbe or microbial DNA in tissue specimens or faecal samples, or the demonstration of L. intracellularis-specific antibodies in sera. The aim of the present study was to evaluate a blocking ELISA in the detection of serum antibodies to L. intracellularis, by comparison to the previously widely used immunofluorescent antibody test (IFAT).MethodsSera were collected from 176 pigs aged 8-12 weeks originating from 24 herds with or without problems with diarrhoea and poor performance in young growing pigs. Sera were analyzed by the blocking ELISA and by IFAT. Bayesian modelling techniques were used to account for the absence of a gold standard test and the results of the blocking ELISA was modelled against the IFAT test with a "2 dependent tests, 2 populations, no gold standard" model.ResultsAt the finally selected cut-off value of percent inhibition (PI) 35, the diagnostic sensitivity of the blocking ELISA was 72% and the diagnostic specificity was 93%. The positive predictive value was 0.82 and the negative predictive value was 0.89, at the observed prevalence of 33.5%.ConclusionThe sensitivity and specificity as evaluated by Bayesian statistic techniques differed from that previously reported. Properties of diagnostic tests may well vary between countries, laboratories and among populations of animals. In the absence of a true gold standard, the importance of validating new methods by appropriate statistical methods and with respect to the target population must be emphasized.