Genetic diversity of equine arteritis virus.

Stadejek, Tomasz; Bj√∂rklund, H; Bascu√±ana, C R; Ciabatti, I.; Scicluna, Maria Teresa; Amaddeo, D.; McCollum, William H.; Autorino, Gian Luca; Timoney, Peter J.; Paton, David; Klingeborn, B.; Bel√°k, S

doi:10.1099/0022-1317-80-3-691

Cited by 54 publications

(61 citation statements)

References 18 publications

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“…The PCR products were run on 2% agarose gels, stained with ethidium bromide, observed under UV light and purified using a PCR purification kit. The sequences of both strands were determined in a MegaBACE TM 1000 with primers CR2 and EAV32, which flank a 519-nt region (8).…”

Section: Reverse-transcription Polymerase Chain Reaction and Sequencingmentioning

confidence: 99%

“…Before the 2010 occurrence of equine viral arteritis in Argentina, the virus had not been involved in respiratory disease, abortion or foal death; the prevalence was restricted to sport-horse breeds and to certain breed lineages (7). Phylogenetic investigations of the virus have focused on the hypervariable region of the GP5 gene, in both European and North American strains (8,9,10). At present, Argentinian EAV strains all belong to the same group (6,11,12).…”

Section: Introductionmentioning

confidence: 99%

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The equine arteritis virus isolate from the 2010 Argentinian outbreak

Metz¹,

Serena²,

Panei³

et al. 2014

Rev. Sci. Tech. OIE

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SummaryA semen sample from a stallion infected during the 2010 equine arteritis virus (EAV) outbreak was received for viral isolation prior to castration of the animal. The virus was identified using a polyclonal antibody immunofluorescence test. Reverse-transcription polymerase chain reaction (RT-PCR) was used to amplify a region of the GP5 gene with primers GL105F and GL673R. The PCR products were purified and sequences of both strands were determined in a MegaBACE ™ 1000 with inner primers CR2 and EAV32. A phylogenetic dataset was built with the previously reported sequences of five strains isolated in Argentina, together with a group of selected sequences obtained from GenBank. The unrooted neighbourjoining tree was constructed using molecular evolutionary genetic analysis (MEGA) and bootstrap analyses were conducted using 1,000 replicate datasets. Evolutionary distances were computed using the maximum composite likelihood method. A NetNGlyc server analysis at the Technical University of Denmark (www.cbs.dtu.dk/services/NetNGlyc/) was used to predict N-glycosylation in GP5 sequences. The phylogenetic analysis revealed that the new strain (GLD-LP-ARG), together with other strains previously isolated, belongs to the European group EU-1 but in a different branch. The new strain shows 99% nucleotide identity with strain A1 and 98.1% with the Belgian strain 08P178. Persistently infected stallions and their cryopreserved semen constitute a reservoir of EAV, which ensures its persistence in the horse population around the world. These findings reinforce the importance of careful monitoring of persistently infected stallions, as well as semen straws, by RT-PCR or test mating, in accordance with national regulations.

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Section: Reverse-transcription Polymerase Chain Reaction and Sequencingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The equine arteritis virus isolate from the 2010 Argentinian outbreak

Metz¹,

Serena²,

Panei³

et al. 2014

Rev. Sci. Tech. OIE

View full text Add to dashboard Cite

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“…1) [19,20]. There is only one known serotype of EAV, but geographically and temporally distinct strains of EAV differ in the severity of the clinical disease they induce and in their abortigenic potential [15,[21][22][23][24][25][26][27][28][29]. Strains of EAV from North America and Europe share at least 85% nucleotide identity; following phylogenetic analysis, these viruses generally segregate into North American and European geographical groups.…”

Section: Etiologymentioning

confidence: 99%

Equine viral arteritis: Current status and prevention

et al. 2008

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“…I), [1,3,7,11,12,14,16]. These viruses, which were propagated in RK-13 cells using established procedures [17], varied in virulence, came from different geographical locations and had been isolated over a period of 30 years.…”

Section: Eav Isolatesmentioning

confidence: 99%

“…RT-PCR has been described as an alternative to virus isolation (VI) for the detection of EAV [2,6,[13][14][15][16], especially for screening semen. In order to improve the reliability and ease of use of RT-PCR, the aim of this work was to compare existing RT-PCR primer sets in order to select a suitable combination for use in conjunction with a fluorescent probe in a one-tube assay.…”

Section: Introductionmentioning

confidence: 99%

Use of an internal standard in a closed one-tube RT-PCR for the detection of equine arteritis virus RNA with fluorescent probes

et al. 2003

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-Routine detection of equine arteritis virus (EAV) can be achieved by virus isolation (VI) in cell culture, or by the amplification of viral genome by molecular methods. To simplify molecular diagnosis, a number of different Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) and RT-nested PCR (RT-nPCR) assays were compared, and a one-tube method was developed and optimised utilizing a fluorogenic probe (TaqMan ® ). An artificial RNA template (Mimic) and associated probe were also constructed to provide in-tube validation of the RT-PCR system. To assess the utility of the RT-PCR TaqMan ® assay, 28 different isolates of EAV representing different genetic groups of American and European strains were tested. Furthermore, the ability of VI and RT-PCR TaqMan ® assay to detect EAV in different biological matrices such as semen, nasal and faecal swabs and blood was compared. All 28 EAV strains were detected by the RT-PCR TaqMan ® assay. The results of TaqMan ® and VI testing were in agreement for 30 of the 33 semen samples and all of the 50 other clinical specimens examined: the RT-PCR TaqMan ® assay detected 18 positive semen samples, three more than VI. In conclusion, the one-tube RT-PCR TaqMan ® assay is a rapid, reliable method for the detection of EAV.equine arteritis virus / RT-PCR / mimic / fluorescent probe / TaqMan ®

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Genetic diversity of equine arteritis virus.

Abstract: Equine arteritis viruses (EAV) from Europe and

Cited by 54 publications

References 18 publications

The equine arteritis virus isolate from the 2010 Argentinian outbreak

The equine arteritis virus isolate from the 2010 Argentinian outbreak

Equine viral arteritis: Current status and prevention

Use of an internal standard in a closed one-tube RT-PCR for the detection of equine arteritis virus RNA with fluorescent probes

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