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

J Virol Methods. 2014 Sep:206:38-41. doi: 10.1016/j.jviromet.2014.05.019. Epub 2014 May 29.

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.

Keywords: Chelonid fibropapilloma-associated herpesvirus; Fibropapillomatosis; Latency; PCR; Sea turtles; Sensitivity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Animals
  • DNA, Viral / analysis*
  • DNA, Viral / genetics
  • Herpesviridae / genetics
  • Herpesviridae / isolation & purification*
  • Herpesviridae Infections / diagnosis
  • Herpesviridae Infections / veterinary*
  • Herpesviridae Infections / virology
  • Molecular Diagnostic Techniques / methods*
  • Papilloma / diagnosis
  • Papilloma / veterinary*
  • Papilloma / virology
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Turtles
  • Veterinary Medicine / methods*

Substances

  • DNA, Viral