Development of in-house Taqman qPCR assay to detect equine herpesvirus-2 in Al-Qadisiyah city
EHV-2 is distributed in horses globally. It is clustered within gamma-herpesvirus subfamily and percavirus genus. EHV-2 infection has two phases: latent and lytic. In the later, EHV-2 mainly associated with respiratory and genital symptoms. However, in the quiescent phase of infection, EHV-2 stay do...
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| Format: | Article |
| Language: | Arabic |
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University of Mosul, College of Veterinary Medicine
2020-05-01
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| Series: | Iraqi Journal of Veterinary Sciences |
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| Online Access: | https://vetmedmosul.com/article_164634_0bcf7eead45c38932c4e5fd354511219.pdf |
| _version_ | 1818605984603439104 |
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| author | Mohammed H. Al-Saadi |
| author_facet | Mohammed H. Al-Saadi |
| author_sort | Mohammed H. Al-Saadi |
| collection | DOAJ |
| description | EHV-2 is distributed in horses globally. It is clustered within gamma-herpesvirus subfamily and percavirus genus. EHV-2 infection has two phases: latent and lytic. In the later, EHV-2 mainly associated with respiratory and genital symptoms. However, in the quiescent phase of infection, EHV-2 stay dormant in the host till viral reactivation. Our previous study has showed that EHV-2 can be harboured by equine tendons, suggesting that leukocytes possibly carrying EHV-2 for the systemic dissemination. So far, numerous PCR protocols have been performed targeting the gB gene. However, this gene is heterogenic. Therefore, there is a need to develop a quantitative diagnostic approach to detect the quiescent EHV-2 strains. To do this, Taqman qPCR assay was developed to quantify the virus. This was performed by targeting a highly conserved gene known as DNA polymerase (DPOL) gene using constructed plasmid as a standard curve calibrator. The obtained results showed an infection frequency of 33% in which the EHV-2 load reached 6647 copies/100 ng DNA whereas the minimum load revealed as 2 copies/100 ng DNA. The median quantification was found as 141 copies/ 100 ng DNA. Establishment of a credited qPCR assay to quantify EHV-2 could be helpful in the control of the disease. |
| first_indexed | 2024-12-16T14:03:39Z |
| format | Article |
| id | doaj.art-5f48378012574d0db0f46dd8b5d3bf9f |
| institution | Directory Open Access Journal |
| issn | 1607-3894 2071-1255 |
| language | Arabic |
| last_indexed | 2024-12-16T14:03:39Z |
| publishDate | 2020-05-01 |
| publisher | University of Mosul, College of Veterinary Medicine |
| record_format | Article |
| series | Iraqi Journal of Veterinary Sciences |
| spelling | doaj.art-5f48378012574d0db0f46dd8b5d3bf9f2022-12-21T22:28:59ZaraUniversity of Mosul, College of Veterinary MedicineIraqi Journal of Veterinary Sciences1607-38942071-12552020-05-0134236537110.33899/ijvs.2019.126076.1229164634Development of in-house Taqman qPCR assay to detect equine herpesvirus-2 in Al-Qadisiyah cityMohammed H. Al-Saadi0Department of Internal and Preventive Medicine, College of Veterinary Medicine, University of Al-Qadisiyah, Al-Qadisiyah, IraqEHV-2 is distributed in horses globally. It is clustered within gamma-herpesvirus subfamily and percavirus genus. EHV-2 infection has two phases: latent and lytic. In the later, EHV-2 mainly associated with respiratory and genital symptoms. However, in the quiescent phase of infection, EHV-2 stay dormant in the host till viral reactivation. Our previous study has showed that EHV-2 can be harboured by equine tendons, suggesting that leukocytes possibly carrying EHV-2 for the systemic dissemination. So far, numerous PCR protocols have been performed targeting the gB gene. However, this gene is heterogenic. Therefore, there is a need to develop a quantitative diagnostic approach to detect the quiescent EHV-2 strains. To do this, Taqman qPCR assay was developed to quantify the virus. This was performed by targeting a highly conserved gene known as DNA polymerase (DPOL) gene using constructed plasmid as a standard curve calibrator. The obtained results showed an infection frequency of 33% in which the EHV-2 load reached 6647 copies/100 ng DNA whereas the minimum load revealed as 2 copies/100 ng DNA. The median quantification was found as 141 copies/ 100 ng DNA. Establishment of a credited qPCR assay to quantify EHV-2 could be helpful in the control of the disease.https://vetmedmosul.com/article_164634_0bcf7eead45c38932c4e5fd354511219.pdfqpcrgamma-herpesvirusehv-2dpol gene |
| spellingShingle | Mohammed H. Al-Saadi Development of in-house Taqman qPCR assay to detect equine herpesvirus-2 in Al-Qadisiyah city Iraqi Journal of Veterinary Sciences qpcr gamma-herpesvirus ehv-2 dpol gene |
| title | Development of in-house Taqman qPCR assay to detect equine herpesvirus-2 in Al-Qadisiyah city |
| title_full | Development of in-house Taqman qPCR assay to detect equine herpesvirus-2 in Al-Qadisiyah city |
| title_fullStr | Development of in-house Taqman qPCR assay to detect equine herpesvirus-2 in Al-Qadisiyah city |
| title_full_unstemmed | Development of in-house Taqman qPCR assay to detect equine herpesvirus-2 in Al-Qadisiyah city |
| title_short | Development of in-house Taqman qPCR assay to detect equine herpesvirus-2 in Al-Qadisiyah city |
| title_sort | development of in house taqman qpcr assay to detect equine herpesvirus 2 in al qadisiyah city |
| topic | qpcr gamma-herpesvirus ehv-2 dpol gene |
| url | https://vetmedmosul.com/article_164634_0bcf7eead45c38932c4e5fd354511219.pdf |
| work_keys_str_mv | AT mohammedhalsaadi developmentofinhousetaqmanqpcrassaytodetectequineherpesvirus2inalqadisiyahcity |