The Experts below are selected from a list of 36 Experts worldwide ranked by ideXlab platform
Bruce G Corney - One of the best experts on this subject based on the ideXlab platform.
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Multiplex real-time PCR for the detection and differentiation of Equid Herpesvirus 1 (EHV-1) and Equid Herpesvirus 4 (EHV-4).
Veterinary microbiology, 2007Co-Authors: Ibrahim S Diallo, Glen Hewitson, Mark A Kelly, Lucia L Wright, Barry J Rodwell, Bruce G CorneyAbstract:A multiplex real-time PCR was designed to detect and differentiate Equid Herpesvirus 1 (EHV-1) and Equid Herpesvirus 4 (EHV-4). The PCR targets the glycoprotein B gene of EHV-1 and EHV-4. Primers and probes were specific to each equine Herpesvirus type and can be used in monoplex or multiplex PCRs, allowing the differentiation of these two closely related members of the Alphaherpesvirinae. The two probes were minor-groove binding probes (MGB) labelled with 6-carboxy-fluorescein (FAM) and VIC for detection of EHV-1 and EHV-4, respectively. Ten EHV-1 isolates, six EHV-1 positive clinical samples, one EHV-1 reference strain (EHV-1.438/77), three EHV-4 positive clinical samples, two EHV-4 isolates and one EHV-4 reference strain (EHV-4 405/76) were included in this study. EHV-1 isolates, clinical samples and the reference strain reacted in the EHV-1 real-time PCR but not in the EHV-4 real-time PCR and similarly EHV-4 clinical samples, isolates and the reference strain were positive in the EHV-4 real-time PCR but not in the EHV-1 real-time PCR. Other Herpesviruses, such as EHV-2, EHV-3 and EHV-5 were all negative when tested using the multiplex real-time PCR. When bacterial pathogens and opportunistic pathogens were tested in the multiplex real-time PCR they did not react with either system. The multiplex PCR was shown to be sensitive and specific and is a useful tool for detection and differentiation of EHV-1 and EHV-4 in a single reaction. A comprehensive equine Herpesvirus disease investigation procedure used in our laboratory is also outlined. This procedure describes the combination of alphaHerpesvirus multiplex real-time PCR along with existing gel-based PCRs described by other authors.
Glen Hewitson - One of the best experts on this subject based on the ideXlab platform.
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equine Herpesvirus infections in yearlings in south east queensland
Archives of Virology, 2008Co-Authors: I S Diallo, Glen Hewitson, Amanda De Jong, Mark A Kelly, Dick J Wright, B G Corney, B J RodwellAbstract:Twelve nasal swabs were collected from yearling horses with respiratory distress and tested for Equid Herpesvirus 1 (EHV-1) and Equid Herpesvirus 4 (EHV-4) by real-time PCR targeting the glycoprotein B gene. All samples were negative for EHV-1; however, 3 were positive for EHV-4. When these samples were tested for EHV-2 and EHV-5 by PCR, all samples were negative for EHV-2 and 11 were positive for EHV-5. All three samples that were positive for EHV-4 were also positive for EHV-5. These three samples gave a limited CPE in ED cells reminiscent of EHV-4 CPE. EHV-4 CPE was obvious after 3 days and was characterised by syncytia. None of the samples produced cytopathic effect (CPE) on African green monkey kidney (Vero) cells or hamster kidney (BSR) cells. Four of the samples, which were positive in the EHV-5 PCR, produced CPE on rabbit kidney (RK13) cells and equine dermis (ED) cells. EHV-5 CPE on both cell lines was slow and was apparent after four 7-day passages. On RK13 cells, the CPE was characteristic of Equid Herpesvirus, with the formation of syncytia. However, in ED cells, the CPE was characterised by ring-shaped syncytia. For the first time, a case of equine respiratory disease involving dual infection with EHV-4 and EHV-5 has been reported in Queensland (Australia). This was shown by simultaneously isolating EHV-4 and EHV-5 from clinical samples. EHV5 was recovered from all samples except one, suggesting that EHV5 was more prevalent in young horses than EHV2.
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Multiplex real-time PCR for the detection and differentiation of Equid Herpesvirus 1 (EHV-1) and Equid Herpesvirus 4 (EHV-4).
Veterinary microbiology, 2007Co-Authors: Ibrahim S Diallo, Glen Hewitson, Mark A Kelly, Lucia L Wright, Barry J Rodwell, Bruce G CorneyAbstract:A multiplex real-time PCR was designed to detect and differentiate Equid Herpesvirus 1 (EHV-1) and Equid Herpesvirus 4 (EHV-4). The PCR targets the glycoprotein B gene of EHV-1 and EHV-4. Primers and probes were specific to each equine Herpesvirus type and can be used in monoplex or multiplex PCRs, allowing the differentiation of these two closely related members of the Alphaherpesvirinae. The two probes were minor-groove binding probes (MGB) labelled with 6-carboxy-fluorescein (FAM) and VIC for detection of EHV-1 and EHV-4, respectively. Ten EHV-1 isolates, six EHV-1 positive clinical samples, one EHV-1 reference strain (EHV-1.438/77), three EHV-4 positive clinical samples, two EHV-4 isolates and one EHV-4 reference strain (EHV-4 405/76) were included in this study. EHV-1 isolates, clinical samples and the reference strain reacted in the EHV-1 real-time PCR but not in the EHV-4 real-time PCR and similarly EHV-4 clinical samples, isolates and the reference strain were positive in the EHV-4 real-time PCR but not in the EHV-1 real-time PCR. Other Herpesviruses, such as EHV-2, EHV-3 and EHV-5 were all negative when tested using the multiplex real-time PCR. When bacterial pathogens and opportunistic pathogens were tested in the multiplex real-time PCR they did not react with either system. The multiplex PCR was shown to be sensitive and specific and is a useful tool for detection and differentiation of EHV-1 and EHV-4 in a single reaction. A comprehensive equine Herpesvirus disease investigation procedure used in our laboratory is also outlined. This procedure describes the combination of alphaHerpesvirus multiplex real-time PCR along with existing gel-based PCRs described by other authors.
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Detection of equine Herpesvirus type 1 using a real-time polymerase chain reaction.
Journal of virological methods, 2005Co-Authors: I S Diallo, Glen Hewitson, B J Rodwell, L.l. Wright, B G CorneyAbstract:Equid Herpesvirus 1 (EHV1) is a major disease of Equids worldwide causing considerable losses to the horse industry. A variety of techniques, including PCR have been used to diagnose EHV1. Some of these PCRs were used in combination with other techniques such as restriction enzyme analysis (REA) or hybridisation, making them cumbersome for routine diagnostic testing and increasing the chances of cross-contamination. Furthermore, they involve the use of suspected carcinogens such as ethidium bromide and ultraviolet light. In this paper, we describe a real-time PCR, which uses minor groove-binding probe (MGB) technology for the diagnosis of EHV1. This technique does not require post-PCR manipulations thereby reducing the risk of cross-contamination. Most importantly, the technique is specific; it was able to differentiate EHV1 from the closely related member of the Alphaherpesvirinae, Equid Herpesvirus 4 (EHV4). It was not reactive with common opportunistic pathogens such as Escherichia coli, Klebsiella oxytoca, Pseudomonas aeruginosa and Enterobacter agglomerans often involved in abortion. Similarly, it did not react with equine pathogens such as Streptococcus equi, Streptococcus equisimilis, Streptococcus zooepidemicus, Taylorella equigenitalis and Rhodococcus equi, which also cause abortion. The results obtained with this technique agreed with results from published PCR methods. The assay was sensitive enough to detect EHV1 sequences in paraffin-embedded tissues and clinical samples. When compared to virus isolation, the test was more sensitive. This test will be useful for the routine diagnosis of EHV1 based on its specificity, sensitivity, ease of performance and rapidity.
J.m. Whalley - One of the best experts on this subject based on the ideXlab platform.
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Epidemiological investigation of Equid Herpesvirus-4 (EHV-4) excretion assessed by nasal swabs taken from thoroughbred foals.
Veterinary microbiology, 1994Co-Authors: James R. Gilkerson, Louisa Jorm, Daria N. Love, Glenda Lawrence, J.m. WhalleyAbstract:Abstract Equid Herpesvirus-4 (EHV-4) was detected in nasal swabs taken from foals using a PCR based test and this information used to study the epidemiology of EHV-4 disease on three Australian Thoroughbred stud farms in NSW in 1992. There was a very high level of agreement (kappa value of 0.84) between the PCR results and virus isolation using cell culture techniques. There was a strong seasonal distribution of EHV-4 shedding. Twenty-five of 26 positive samples were collected in January and March with the remaining positive sample collected in February. Foals with clinical signs of upper respiratory tract infection per se were no more likely to be shedders of EHV-4 (odds ratio [OR] 1.4, 95% confidence limits [CL] 0.5–3.8). However, EHV-4 was more likely to be isolated from foals exhibiting copious serous or mucopurulent nasal discharge than those with no clinical signs (OR 4.6, 95% CL 1.1–19.0 and OR 2.5, 95% CL 0.8–8.0, respectively). The month of the year was more important than weaning or age as a risk factor for excretion of EHV-4. Male foals and those with a history of respiratory disease that had required veterinary treatment were more likely to shed EHV-4.
Mark A Kelly - One of the best experts on this subject based on the ideXlab platform.
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equine Herpesvirus infections in yearlings in south east queensland
Archives of Virology, 2008Co-Authors: I S Diallo, Glen Hewitson, Amanda De Jong, Mark A Kelly, Dick J Wright, B G Corney, B J RodwellAbstract:Twelve nasal swabs were collected from yearling horses with respiratory distress and tested for Equid Herpesvirus 1 (EHV-1) and Equid Herpesvirus 4 (EHV-4) by real-time PCR targeting the glycoprotein B gene. All samples were negative for EHV-1; however, 3 were positive for EHV-4. When these samples were tested for EHV-2 and EHV-5 by PCR, all samples were negative for EHV-2 and 11 were positive for EHV-5. All three samples that were positive for EHV-4 were also positive for EHV-5. These three samples gave a limited CPE in ED cells reminiscent of EHV-4 CPE. EHV-4 CPE was obvious after 3 days and was characterised by syncytia. None of the samples produced cytopathic effect (CPE) on African green monkey kidney (Vero) cells or hamster kidney (BSR) cells. Four of the samples, which were positive in the EHV-5 PCR, produced CPE on rabbit kidney (RK13) cells and equine dermis (ED) cells. EHV-5 CPE on both cell lines was slow and was apparent after four 7-day passages. On RK13 cells, the CPE was characteristic of Equid Herpesvirus, with the formation of syncytia. However, in ED cells, the CPE was characterised by ring-shaped syncytia. For the first time, a case of equine respiratory disease involving dual infection with EHV-4 and EHV-5 has been reported in Queensland (Australia). This was shown by simultaneously isolating EHV-4 and EHV-5 from clinical samples. EHV5 was recovered from all samples except one, suggesting that EHV5 was more prevalent in young horses than EHV2.
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Multiplex real-time PCR for the detection and differentiation of Equid Herpesvirus 1 (EHV-1) and Equid Herpesvirus 4 (EHV-4).
Veterinary microbiology, 2007Co-Authors: Ibrahim S Diallo, Glen Hewitson, Mark A Kelly, Lucia L Wright, Barry J Rodwell, Bruce G CorneyAbstract:A multiplex real-time PCR was designed to detect and differentiate Equid Herpesvirus 1 (EHV-1) and Equid Herpesvirus 4 (EHV-4). The PCR targets the glycoprotein B gene of EHV-1 and EHV-4. Primers and probes were specific to each equine Herpesvirus type and can be used in monoplex or multiplex PCRs, allowing the differentiation of these two closely related members of the Alphaherpesvirinae. The two probes were minor-groove binding probes (MGB) labelled with 6-carboxy-fluorescein (FAM) and VIC for detection of EHV-1 and EHV-4, respectively. Ten EHV-1 isolates, six EHV-1 positive clinical samples, one EHV-1 reference strain (EHV-1.438/77), three EHV-4 positive clinical samples, two EHV-4 isolates and one EHV-4 reference strain (EHV-4 405/76) were included in this study. EHV-1 isolates, clinical samples and the reference strain reacted in the EHV-1 real-time PCR but not in the EHV-4 real-time PCR and similarly EHV-4 clinical samples, isolates and the reference strain were positive in the EHV-4 real-time PCR but not in the EHV-1 real-time PCR. Other Herpesviruses, such as EHV-2, EHV-3 and EHV-5 were all negative when tested using the multiplex real-time PCR. When bacterial pathogens and opportunistic pathogens were tested in the multiplex real-time PCR they did not react with either system. The multiplex PCR was shown to be sensitive and specific and is a useful tool for detection and differentiation of EHV-1 and EHV-4 in a single reaction. A comprehensive equine Herpesvirus disease investigation procedure used in our laboratory is also outlined. This procedure describes the combination of alphaHerpesvirus multiplex real-time PCR along with existing gel-based PCRs described by other authors.
Ibrahim S Diallo - One of the best experts on this subject based on the ideXlab platform.
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Multiplex real-time PCR for the detection and differentiation of Equid Herpesvirus 1 (EHV-1) and Equid Herpesvirus 4 (EHV-4).
Veterinary microbiology, 2007Co-Authors: Ibrahim S Diallo, Glen Hewitson, Mark A Kelly, Lucia L Wright, Barry J Rodwell, Bruce G CorneyAbstract:A multiplex real-time PCR was designed to detect and differentiate Equid Herpesvirus 1 (EHV-1) and Equid Herpesvirus 4 (EHV-4). The PCR targets the glycoprotein B gene of EHV-1 and EHV-4. Primers and probes were specific to each equine Herpesvirus type and can be used in monoplex or multiplex PCRs, allowing the differentiation of these two closely related members of the Alphaherpesvirinae. The two probes were minor-groove binding probes (MGB) labelled with 6-carboxy-fluorescein (FAM) and VIC for detection of EHV-1 and EHV-4, respectively. Ten EHV-1 isolates, six EHV-1 positive clinical samples, one EHV-1 reference strain (EHV-1.438/77), three EHV-4 positive clinical samples, two EHV-4 isolates and one EHV-4 reference strain (EHV-4 405/76) were included in this study. EHV-1 isolates, clinical samples and the reference strain reacted in the EHV-1 real-time PCR but not in the EHV-4 real-time PCR and similarly EHV-4 clinical samples, isolates and the reference strain were positive in the EHV-4 real-time PCR but not in the EHV-1 real-time PCR. Other Herpesviruses, such as EHV-2, EHV-3 and EHV-5 were all negative when tested using the multiplex real-time PCR. When bacterial pathogens and opportunistic pathogens were tested in the multiplex real-time PCR they did not react with either system. The multiplex PCR was shown to be sensitive and specific and is a useful tool for detection and differentiation of EHV-1 and EHV-4 in a single reaction. A comprehensive equine Herpesvirus disease investigation procedure used in our laboratory is also outlined. This procedure describes the combination of alphaHerpesvirus multiplex real-time PCR along with existing gel-based PCRs described by other authors.