Validation of a sensitive PCR assay for the detection of Chelonid fibropapilloma-associated herpesvirus in latent turtle infections

The Chelonid fibropapilloma-associated herpesvirus (CFPHV) is hypothesized to be the causative agent of fibropapillomatosis, a neoplastic disease in sea turtles, given its consistent detection by PCR in fibropapilloma tumours. CFPHV has also been detected recently by PCR in tissue samples from clini...

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Main Authors: Alfaro-Núñez, A., Gilbert, Thomas
Format: Journal Article
Published: Elsevier BV 2014
Subjects:
Online Access:http://hdl.handle.net/20.500.11937/12708
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author Alfaro-Núñez, A.
Gilbert, Thomas
author_facet Alfaro-Núñez, A.
Gilbert, Thomas
author_sort Alfaro-Núñez, A.
building Curtin Institutional Repository
collection Online Access
description The Chelonid fibropapilloma-associated herpesvirus (CFPHV) is hypothesized to be the causative agent of fibropapillomatosis, a neoplastic disease in sea turtles, given its consistent detection by PCR in fibropapilloma tumours. CFPHV has also been detected recently by PCR in tissue samples from clinically healthy (non exhibiting fibropapilloma tumours) turtles, thus representing presumably latent infections of the pathogen. Given that template copy numbers of viruses in latent infections can be very low, extremely sensitive PCR assays are needed to optimize detection efficiency. In this study, efficiency of several PCR assays designed for CFPHV detection is explored and compared to a method published previously. The results show that adoption of a triplet set of singleplex PCR assays outperforms other methods, with an approximately 3-fold increase in detection success in comparison to the standard assay. Thus, a new assay for the detection of CFPHV DNA markers is presented, and adoption of its methodology is recommended in future CFPHV screens among sea turtles.
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spelling curtin-20.500.11937-127082017-09-13T15:01:03Z Validation of a sensitive PCR assay for the detection of Chelonid fibropapilloma-associated herpesvirus in latent turtle infections Alfaro-Núñez, A. Gilbert, Thomas Latency Chelonid fibropapilloma-associated herpesvirus Sensitivity Sea turtles Fibropapillomatosis PCR The Chelonid fibropapilloma-associated herpesvirus (CFPHV) is hypothesized to be the causative agent of fibropapillomatosis, a neoplastic disease in sea turtles, given its consistent detection by PCR in fibropapilloma tumours. CFPHV has also been detected recently by PCR in tissue samples from clinically healthy (non exhibiting fibropapilloma tumours) turtles, thus representing presumably latent infections of the pathogen. Given that template copy numbers of viruses in latent infections can be very low, extremely sensitive PCR assays are needed to optimize detection efficiency. In this study, efficiency of several PCR assays designed for CFPHV detection is explored and compared to a method published previously. The results show that adoption of a triplet set of singleplex PCR assays outperforms other methods, with an approximately 3-fold increase in detection success in comparison to the standard assay. Thus, a new assay for the detection of CFPHV DNA markers is presented, and adoption of its methodology is recommended in future CFPHV screens among sea turtles. 2014 Journal Article http://hdl.handle.net/20.500.11937/12708 10.1016/j.jviromet.2014.05.019 Elsevier BV restricted
spellingShingle Latency
Chelonid fibropapilloma-associated herpesvirus
Sensitivity
Sea turtles
Fibropapillomatosis
PCR
Alfaro-Núñez, A.
Gilbert, Thomas
Validation of a sensitive PCR assay for the detection of Chelonid fibropapilloma-associated herpesvirus in latent turtle infections
title Validation of a sensitive PCR assay for the detection of Chelonid fibropapilloma-associated herpesvirus in latent turtle infections
title_full Validation of a sensitive PCR assay for the detection of Chelonid fibropapilloma-associated herpesvirus in latent turtle infections
title_fullStr Validation of a sensitive PCR assay for the detection of Chelonid fibropapilloma-associated herpesvirus in latent turtle infections
title_full_unstemmed Validation of a sensitive PCR assay for the detection of Chelonid fibropapilloma-associated herpesvirus in latent turtle infections
title_short Validation of a sensitive PCR assay for the detection of Chelonid fibropapilloma-associated herpesvirus in latent turtle infections
title_sort validation of a sensitive pcr assay for the detection of chelonid fibropapilloma-associated herpesvirus in latent turtle infections
topic Latency
Chelonid fibropapilloma-associated herpesvirus
Sensitivity
Sea turtles
Fibropapillomatosis
PCR
url http://hdl.handle.net/20.500.11937/12708