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Diseases of Aquatic Organisms

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DAO 129:193-198 (2018)  -  DOI: https://doi.org/10.3354/dao03242

Partial validation of a TaqMan real-time quantitative PCR assay for the detection of Panulirus argus virus 1

Abigail S. Clark1,5, Donald C. Behringer1,2, Jessica Moss Small3, Thomas B. Waltzek4,*

1Fisheries and Aquatic Sciences, University of Florida, Gainesville, FL 32653, USA
2Emerging Pathogens Institute, University of Florida, Gainesville, FL 32611, USA
3Aquaculture Genetics and Breeding Technology Center, Virginia Institute of Marine Science, Gloucester Point, VA 23062, USA
4Department of Infectious Diseases and Immunology, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA
5Present address: The Elizabeth Moore International Center for Coral Reef Research and Restoration, Mote Marine Laboratory, 24244 Overseas Highway, Summerland Key, FL 33042, USA
*Corresponding author:

ABSTRACT: The Caribbean spiny lobster Panulirus argus supports important fisheries throughout the greater Caribbean and is also the only known host for the pathogenic virus Panulirus argus virus 1 (PaV1). While discovered nearly 2 decades ago, gaps still exist in our knowledge of PaV1, such as the dose required to establish infection and its viability outside of the host. To help answer such questions and to enhance diagnostic capabilities, we developed a TaqMan real-time quantitative polymerase chain reaction (qPCR) assay for PaV1. Of the advantages offered by qPCR, one of the most important benefits is its ability to accurately quantify viral DNA copies in a clinical sample. The qPCR assay was found to be efficient (mean ± SD: 99.19 ± 4.67%) and sensitive, detecting as few as 10 copies of PaV1 plasmid DNA. Its diagnostic sensitivity and specificity determined using a set of 165 lobster samples (138 from Florida, USA, and 27 from across the Caribbean) were 100 and 84%, respectively. The qPCR assay should thus prove useful as a research tool and for detecting and quantifying PaV1 infection severity in Caribbean spiny lobsters.


KEY WORDS: Panulirus argus virus 1 · Quantitative PCR · Lobster · Diagnostics · Sensitivity · Specificity


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Cite this article as: Clark AS, Behringer DC, Moss Small J, Waltzek TB (2018) Partial validation of a TaqMan real-time quantitative PCR assay for the detection of Panulirus argus virus 1. Dis Aquat Org 129:193-198. https://doi.org/10.3354/dao03242

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