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Characterization of Ovine Herpesviru...
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Alhajri, Salim Mohamed.
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Characterization of Ovine Herpesvirus 2 Putative Glycoprotein Ov8.
紀錄類型:
書目-電子資源 : Monograph/item
正題名/作者:
Characterization of Ovine Herpesvirus 2 Putative Glycoprotein Ov8./
作者:
Alhajri, Salim Mohamed.
出版者:
Ann Arbor : ProQuest Dissertations & Theses, : 2017,
面頁冊數:
68 p.
附註:
Source: Dissertation Abstracts International, Volume: 79-04(E), Section: B.
Contained By:
Dissertation Abstracts International79-04B(E).
標題:
Veterinary science. -
電子資源:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10602639
ISBN:
9780355364460
Characterization of Ovine Herpesvirus 2 Putative Glycoprotein Ov8.
Alhajri, Salim Mohamed.
Characterization of Ovine Herpesvirus 2 Putative Glycoprotein Ov8.
- Ann Arbor : ProQuest Dissertations & Theses, 2017 - 68 p.
Source: Dissertation Abstracts International, Volume: 79-04(E), Section: B.
Thesis (Ph.D.)--Washington State University, 2017.
Malignant catarrhal fever (MCF) is a lymphoproliferative usually fatal disease of ungulates caused by several gammaherpesviruses in the genus Macavirus including alcelaphine herpesvirus 1 (AlHV-1) and ovine herpesvirus 2 (OvHV-2). There is no treatment or vaccine for the disease. MCF is diagnosed by clinical history, histopathology, polymerase chain reaction and serology. However, all current serological assays utilize AlHV-1 antigens thus none are specific for detecting anti-OvHV2 antibodies. The lack of cell culture system to propagate OvHV-2 has hindered studies of OvHV-2 biology including its entry mechanism and in making of an OvHV-2 specific antibody detection assay. As an alternative method to study OvHV-2 glycoproteins responsible for membrane fusion as a part of the entry mechanism, we developed a virus-free cell-to-cell membrane fusion assay to identify the minimum required OvHV-2 glycoproteins to induce membrane fusion. OvHV-2 glycoproteins gB, gH and gL could induce membrane fusion together but not individually. Cell-cell membrane fusion was detected by both fluorescence microscopy and reporter gene expression. Additionally, open reading frame Ov8, unique to OvHV-2, was found to encode a transmembrane glycoprotein that can significantly enhance membrane fusion. These results are described in chapter 1. To overcome the lack of cell culture to grow OvHV-2, a recombinant fusion protein containing part of the OvHV-2-specific Ov8 glycoprotein was expressed and evaluated in an enzyme linked immunosorbent assay (ELISA). Using sheep serum samples, the assay showed relatively 100% sensitivity, 98.97% specificity, 99.07% positive predictive value, and 100% negative predictive value. The assay also showed very good agreement with a competitive inhibition (CI)-ELISA (kappa = 0.990 and 95% CI = 0.971--1.000) that detects an epitope conserved among all MCF viruses. Using samples from bison with MCF, both the CI-ELISA and Ov8 ELISA showed 58.82% sensitivity and100% specificity. Importantly, 31.5% of samples not detected by CI-ELISA were detected in Ov8 ELISA. These results show that Ov8 ELISA has potential as an OvHV-2 specific diagnostic assay.
ISBN: 9780355364460Subjects--Topical Terms:
3172798
Veterinary science.
Characterization of Ovine Herpesvirus 2 Putative Glycoprotein Ov8.
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Malignant catarrhal fever (MCF) is a lymphoproliferative usually fatal disease of ungulates caused by several gammaherpesviruses in the genus Macavirus including alcelaphine herpesvirus 1 (AlHV-1) and ovine herpesvirus 2 (OvHV-2). There is no treatment or vaccine for the disease. MCF is diagnosed by clinical history, histopathology, polymerase chain reaction and serology. However, all current serological assays utilize AlHV-1 antigens thus none are specific for detecting anti-OvHV2 antibodies. The lack of cell culture system to propagate OvHV-2 has hindered studies of OvHV-2 biology including its entry mechanism and in making of an OvHV-2 specific antibody detection assay. As an alternative method to study OvHV-2 glycoproteins responsible for membrane fusion as a part of the entry mechanism, we developed a virus-free cell-to-cell membrane fusion assay to identify the minimum required OvHV-2 glycoproteins to induce membrane fusion. OvHV-2 glycoproteins gB, gH and gL could induce membrane fusion together but not individually. Cell-cell membrane fusion was detected by both fluorescence microscopy and reporter gene expression. Additionally, open reading frame Ov8, unique to OvHV-2, was found to encode a transmembrane glycoprotein that can significantly enhance membrane fusion. These results are described in chapter 1. To overcome the lack of cell culture to grow OvHV-2, a recombinant fusion protein containing part of the OvHV-2-specific Ov8 glycoprotein was expressed and evaluated in an enzyme linked immunosorbent assay (ELISA). Using sheep serum samples, the assay showed relatively 100% sensitivity, 98.97% specificity, 99.07% positive predictive value, and 100% negative predictive value. The assay also showed very good agreement with a competitive inhibition (CI)-ELISA (kappa = 0.990 and 95% CI = 0.971--1.000) that detects an epitope conserved among all MCF viruses. Using samples from bison with MCF, both the CI-ELISA and Ov8 ELISA showed 58.82% sensitivity and100% specificity. Importantly, 31.5% of samples not detected by CI-ELISA were detected in Ov8 ELISA. These results show that Ov8 ELISA has potential as an OvHV-2 specific diagnostic assay.
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