Identification of equine herpesviruses 1 and 4 by polymerase chain reaction

Varrasso, Annalisa; Dynon, Kemperly; Ficorilli, Nino; Hartley, Carol A.; Studdert, M. J.; Drummer, Heidi E.

doi:10.1111/j.1751-0813.2001.tb10751.x

Cited by 51 publications

(44 citation statements)

References 23 publications

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“…The sensitivity of the PCR used was higher when compared with similar data presented by Varrasso et al (2001) who found that their glycoprotein H gene-based PCR was able to detect 1220 and 7280 plasmid EHV1 and EHV4 DNA copies, respectively. The authors increased the sensitivity of the assay by adding a second amplification cycle using nested primers.…”

Section: Discussionmentioning

confidence: 69%

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

Ploszay¹,

Rola²,

Larska³

et al. 2013

Polish Journal of Veterinary Sciences

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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 10 3 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.

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Section: Discussionmentioning

confidence: 69%

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

Ploszay¹,

Rola²,

Larska³

et al. 2013

Polish Journal of Veterinary Sciences

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“…As amostras negativas para o vírus da raiva foram enviadas ao Laboratório de Patologia Molecular da Escola de Veterinária da UFMG para o diagnóstico através de PCR específicos para EHV-1 e EHV-4 (Varrasso et al, 2001), SHV-1 (Fonseca Jr. et al, 2011) ou RT-PCR para as seguintes arboviroses: SLEV (Ré et al, 2008), WNV, EEEV (Johnson et al, 2003), WEEV e VEEV (Eshoo et al, 2007). Para a extração do DNA/RNA total, as amostras de fragmentos de SNC (córtex, cerebelo, hipocampo, tálamo e medula) foram suspensas em solução salina tamponada a 20% (p/v) e livre de DNA e RNA, homogeneizadas e centrifugadas a 2.000 x g, a 4 o C, por 30 minutos.…”

Section: Methodsunclassified

“…A segunda reação dos semi-nested PCR (EHV-1 e 4) foi feita utilizando-se a mesma concentração dos reagentes da primeira reação e adicionando-se como amplicon, 1μL da primeira reação. Os oligonucleotídeos iniciadores específicos utilizados para detecção dos EHV-1, EHV-4 e SHV-1 foram desenhados e descritos previamente (Varrasso et al, 2001;Fonseca Jr. et al, 2011). Para assegurar a qualidade das amostras de DNA, foi realizada amplificação de parte do gene do fator de crescimento semelhante à insulina 1 (IGF-1), altamente conservado em mamíferos (Mikawa et al, 1995), sob condições previamente descritas (Costa et al, 2009).…”

Section: Methodsunclassified

“…Dentre as 170 amostras de SNC negativas para o vírus da raiva, provenientes de 82 municípios abrangendo todas as 12 mesorregiões de Minas Gerais, 20 (11,8%) foram positivas para EHV-1 através da técnica de semi-nested PCR (Varrasso et al, 2001). Não houve detecção do EHV-4 no DNA oriundo do SNC de nenhum dos equinos analisados, o que indica baixa frequência desse vírus no Estado de Minas Gerais, uma vez que a metodologia empregada apresenta elevada sensibilidade e especificidade (Varrasso et al 2001).…”

Section: Methodsunclassified

“…Não houve detecção do EHV-4 no DNA oriundo do SNC de nenhum dos equinos analisados, o que indica baixa frequência desse vírus no Estado de Minas Gerais, uma vez que a metodologia empregada apresenta elevada sensibilidade e especificidade (Varrasso et al 2001).…”

Section: Methodsunclassified

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Diagnóstico etiológico de enfermidades do sistema nervoso central de equinos no Estado de Minas Gerais, Brasil

Costa

Rosa

Oliveira³

et al. 2015

Arq. Bras. Med. Vet. Zootec.

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 RESUMOO Brasil possui o quarto maior rebanho equino do mundo, e o Estado de Minas Gerais detém o maior número de equinos do país. Portanto, um diagnóstico preciso das doenças neurológicas dos equinos é prioridade no estado. Sendo assim, o objetivo deste estudo foi identificar, utilizando a Reação em Cadeia pela Polimerase (PCR), os agentes infecciosos responsáveis por enfermidades que afetam o sistema nervoso central (SNC) de equinos. De janeiro de 2009 a janeiro de 2011, foi realizado um levantamento dos casos de encefalites e encefalomielites em equinos no Estado de Minas Gerais, utilizando-se amostras de SNC de equinos que morreram com sinais neurológicos. Das 217 amostras de SNC, 47 (21,7%) foram positivas para o vírus da raiva pelo método de imunofluorescência indireta e inoculação em camundongos. Nas 170 amostras negativas para o vírus da raiva, o herpes-vírus equino-1 (EHV-1) foi diagnosticado em 20 (11,8%) e o herpes-vírus suíno-1 (SHV-1), em uma amostra por meio de PCR, e o vírus encefalite de Saint Louis (SLEV), em outra amostra, através de transcrição reversa (RT) e PCR (RT-PCR). Constatou-se que o vírus da raiva é o principal agente causador de encefalite em equinos, apesar do crescente número de casos de encefalomielite associados ao EHV-1 no Estado de Minas Gerais. Palavras-chave: cavalo, diagnóstico, encefalite, Brasil ABSTRACT Brazil has the fourth largest equine herd in the world and the State of Minas Gerais has the largest equine population in the country. Therefore, an accurate diagnosis of cases of neurologic diseases is a priority in Minas Gerais. The aim of this study was to identify by Polymerase Chain Reaction (PCR) infectious agents associated with neurological disease in the central nervous system (CNS) of horses. A survey of encephalitis and encephalomyelitis in horses in Minas Gerais State was performed on samples of CNS from horses that died with neurological signs from January 2009 to January 2011. Forty seven CNS samples from 217 (21.7%) horses were positive for rabies virus by the indirect immunofluorescence assay and mouse inoculation. Among the 170 samples that were negative for rabies, EHV-1 was detected in 20 (11.8%) and the swine herpesvirus-1 (SHV-1) was detected in one sample by PCR, and the Saint Louis encephalitis virus (SLEV) was identified in another sample by reverse transcription (RT) and PCR (RT-PCR). Rabies virus is the most common causative agent of encephalitis in horses, despite the increasing number of cases of encephalitis associated with EHV-1 in the State of Minas Gerais.

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Neospora spp. infection associated with equine abortion and/or stillbirth rate

et al. 2008

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Identification of equine herpesviruses 1 and 4 by polymerase chain reaction

Abstract: Individual single round and second round (seminested) PCRs and a seminested multiplex PCR were developed that enabled reliable, rapid detection of EHV1 and EHV4 in aborted foetal tissues and nasopharyngeal swab samples.

Cited by 51 publications

References 23 publications

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

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

Diagnóstico etiológico de enfermidades do sistema nervoso central de equinos no Estado de Minas Gerais, Brasil

Neospora spp. infection associated with equine abortion and/or stillbirth rate

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