Development of Quantitative Real Time PCR Assay For The Detection of Chelonid Fibropapilloma-Associated Herpesviruses
| Main Authors: | Nainu, Firzan, Ariel, Ellen, Burgess, Graham W. |
|---|---|
| Format: | Lainnya |
| Bahasa: | eng |
| Terbitan: |
, 2014
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| Subjects: | |
| Online Access: |
http://repository.unhas.ac.id/handle/123456789/8406 |
Daftar Isi:
- Fibropapillomatosis (FP) is a transmissible neoplastic disease characterised by single to multiple external and/or internal benign tumours that are recognised to cause morbidity and mortality in marine turtles worldwide. The aetiology of FP remains inconclusive but, the association of chelonid fibropapilloma-associated herpesvirus (CFPHV) with the development of FP has been widely acknowledged. This study addresses the need for efficient and sensitive diagnostic tools in the detection of CFPHV. The objective of this study was to develop a Syto??9based qPCR assay for the detection and quantification of CFPHV in twenty-three fibropapilloma samples collected from Australian marine turtles. The qPCR method used in this study was able to detect CFPHV ranging from 3.05 ?? 101 to 3.05 ?? 109 copies per reaction with the efficiency of reaction 0.97. High resolution melting (HRM) analysis revealed five possible variants of CFPHV circulating in Australian waters, which is to a certain degree, associated with the geographical collection sites of their respective hosts. The high sensitivity and efficiency of the Syto??9-based qPCR assay, combined with the HRM analysis, make this method suitable for the diagnostic purposes of CFPHV.