First report on equine herpesvirus type 4 isolation in Poland – evaluation of diagnostic tools

• Autorzy: Ploszay G. , Rola J. , Larska M. , Zmudzinski J. F.
• Języki publikacji: EN

Abstrakty

EN

Upper respiratory tract infections are still a serious problem in breeding and racing horses. The most common virological factors are EHV1 and EHV4, which are both a major cause of secondary infections. High EHV4 seroprevalence in Polish horses indicates a high transmission rate of this pathogen among horses and increases the need for proper diagnostics. The aim of this study was to develop a reliable laboratory diagnostic scheme for upper respiratory tract infections and to describe the first isolation of EHV4 in Poland. Twenty one nasal swabs collected from young horses under the age of 2 years showing clinical signs of equine rhinopneumonitis were tested with duplex PCR for simultaneous detection and differentiation between EHV1/EHV4. Positive samples were then subjected to virus isolation in Vero cells. Additionally, real-time PCR was developed which allowed viral copy numbers to be quantified and enabled defining that a DNA load below 10³ copies per 1 ml of the sample reflected latent infection or decline of the disease. However, the sensitivity of traditional PCR proved to be sufficient in the diagnostic of the lytic infections and allowed identification of 10 EHV4 infected horses from which 3 strains were successfully isolated in cell culture. Another four EHV4 positive results were obtained by real-time PCR; but, a high Ct (threshold cycle) and a low virus DNA copy number suggested a latent infection. This report describes the first successful isolation of EHV4 from Polish horses.

• Wydawca: -
• Czasopismo: Polish Journal of Veterinary Sciences
• Rocznik: 2013
• Tom: 16
• Numer: 3
• Opis fizyczny: p.493-500,fig.,ref.

Twórcy

autor

Ploszay G.

• Department of Virology, National Veterinary Research Institute, Al. Partyzantów 57, 24-100 Pulawy, Poland

autor

Rola J.

• Department of Virology, National Veterinary Research Institute, Al. Partyzantów 57, 24-100 Pulawy, Poland

autor

Larska M.

• Department of Virology, National Veterinary Research Institute, Al. Partyzantów 57, 24-100 Pulawy, Poland

autor

Zmudzinski J. F.

• Department of Virology, National Veterinary Research Institute, Al. Partyzantów 57, 24-100 Pulawy, Poland

Bibliografia

• Allen GP, Bryans JT (1986) Molecular epizootiology, pathogenesis and prophylaxis of equine herpesvirus-1 infections. Prog Vet Microbiol Immunol 2: 78-144.
• Blunden AS, Smith KC, Binns MM, Zhang L, Gower SM, Mumford JA (1995) Replication of Equid Herpesvirus 4 in Endothelial Cells and Synovia of a Field Case of Viral Pneumonia and Synovitis in a Foal. J Comp Pathol 112: 133-140.
• Chowdhury SI, Buhk HJ, Ludwig H, Hammerschmidt W (1990) Genomic termini of equine herpesvirus 1. J Virol 64: 873-880.
• Diallo IS, Hewitson G, Wright LL, Kelly MA, Rodwell BJ, Corney BG (2007) Multiplex real-time PCR for the detection and differentiation of equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4). Vet Microbiol 123: 93-103.
• Dynon K, Black WD, Ficorilli N, Hartley CA, Studdert MJ (2007) Detection of viruses in nasal swab samples from horses with acute, febrile, respiratory disease using virus isolation, polymerase chain reaction and serology. Aust Vet J 85: 46-50.
• Gilkerson J, Jorm LR, Love DN, Lawrence GL, Whalley JM (1994) Epidemiological investigation of equid herpesvirus-4 (EHV-4) excretion assessed by nasal swabs taken from thoroughbred foals. Vet Microbiol 39: 275-283.
• Gradzki Z, Boguta L (2009a) Seroprevalence of EHV1 and EHV4 in the horse population of the southeastern part of Poland. Med Weter 65: 188-193.
• Gradzki Z, Boguta L (2009b) Use of PCR and cell culture method in the diagnosis and differentiation of EHV1 and EHV4 infections in horses. Med Weter 65: 546-551.
• Heldens JG, Hannant D, Cullinane AA, Prendergast MJ, Mumford JA, Nelly M, Kydd JH, Weststrate MW, van den Hoven R (2001) Clinical and virological evaluation of the efficacy of an inactivated EHV1 and EHV4 whole virus vaccine (Duvaxyn EHV1,4). Vaccination/challenge experiments in foals and pregnant mares. Vaccine 19: 4307-4317.
• Kirisawa R, Endo A, Iwai H, Kawakami Y (1993) Detection and identification of equine herpesvirus-1 and -4 by polymerase chain reaction. Vet Microbiol 36: 57-67.
• Kirisawa R, Ohmori H, Iwai H, Kawakami Y (1993) The genomic diversity among equine herpesvirus-1 strains isolated in Japan. Arch Virol 129: 11-22.
• Maeda K, Kai K, Matsumura T (2005) Genomic Diversity among Equine Herpesvirus-4 Field Isolates. J Vet Med Sci 67: 555-561.
• McCann SHE, Mumford JA, Binns MM (1995) Development of PCR assays to detect genetic variation amongst equine herpesvirus-1 isolates as an aid to epidemiological investgation. J Virol Methods 52: 183-194.
• Patel JR, Heldens J (2005) Equine herpesviruses 1 (EHV-1) and 4 (EHV-4) – epidemiology, disease and immunoprophylaxis: A brief review. Vet J 170: 14-23.
• Pusterla N, Leutenegger CM, Wilson WD, Watson JL, Ferraro GL, Madigan JE (2005) Equine Herpesvirus-4 Kinetics in Peripheral Blood Leukocytes and Nasopharyngeal Secretions in Foals Using Quantitative Real-Time TaqMan PCR. J Vet Diagn Invest 17: 578-581.
• Rola J (2000) Restriction profiles of Polish isolates of equine herpes virus type 1. Med Weter 56: 813-815.
• Studdert MJ (1983) Restriction endonuclease DNA fingerprinting of respiratory, foetal and perinatal foal isolates of equine herpesvirus type 1. Arch Virol 77: 249-258.
• Varrasso A, Dynon K, Ficorilli N, Hartley CA, Studdert MJ, Drummer HE (2001) Identification of equine herpesviruses 1 and 4 by polymerase chain reaction. Aust Vet J 79: 563-569.
• Wang L, Raidal SL, Pizzirani A, Wilcox GE (2007) Detection of respiratory herpesviruses in foals and adult horses determined by nested multiplex PCR. Vet Microbiol 121: 18-28.
• Wood JL, Newton JR, Chanter N, Mumford JA (2005) Association between respiratory disease and bacterial and viral iffections in British racehorses. J Clin Microbiol 43: 120-126.