title: Validation of a sensitive PCR assay for the detection of Chelonid fibropapilloma-associated herpesvirus in latent turtle infections.
abstract: 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.
authors.affiliation: Section for Evolutionary Genomics, Centre for GeoGenetics, Natural History Museum of Denmark, Øster Voldgade 5-7, 1350 Copenhagen K, Denmark. Electronic address: alonzoalfaro@gmail.com.
authors.forename: Alonzo
authors.initials: A
authors.lastname: Alfaro-Núñez
authors.affiliation: Section for Evolutionary Genomics, Centre for GeoGenetics, Natural History Museum of Denmark, Øster Voldgade 5-7, 1350 Copenhagen K, Denmark; Trace and Environmental DNA Laboratory, School of Environment and Agriculture, Curtin University, Perth, Western Australia 6845, Australia.
authors.forename: M Thomas P
authors.initials: MT
authors.lastname: Gilbert
mesh_terms: D000818:Animals; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D010212:Papilloma / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014426:Turtles; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D004279:DNA, Viral
keywords: Chelonid fibropapilloma-associated herpesvirus; Fibropapillomatosis; Latency; PCR; Sea turtles; Sensitivity