Abstract
BackgroundThe myxozoan Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), is responsible for considerable losses in farmed and wild fish populations in Europe and North America. Recently, T. bryosalmonae was detected in many European countries, and strategy to control the disease in the wild and farmed fish population is yet to be developed. Recombinase polymerase amplification (RPA) is a novel isothermal nucleic acid amplification technology that does not require any thermal cycling, and lateral flow dipstick (LFD) is a rapid, cost-effective, and easy-to-handle assay that enables stable detection.ResultsIn this study, we developed and optimized a rapid and sensitive RPA assay combined with an LFD for the detection of T. bryosalmonae. The PKD-RPA assay was specific to T. bryosalmonae, as no cross-reaction or false positive signals were observed with any of the other tested DNAs. The developed PKD-RPA assay was ten times more sensitive than an existing diagnostic polymerase chain reaction (PCR) assay for this parasite. The estimated time to perform PKD-RPA assay is 25 min compared to 4 h for PKD-PCR assay.ConclusionsA novel PKD-RPA assay for the detection of T. bryosalmonae was developed. The assay offers considerable advantages including speed, sensitivity, specificity and visual detection. Applying the PKD-RPA assay combined with an LFD enhances the surveillance and early detection of T. bryosalmonae in salmonids.
Highlights
The myxozoan Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), is responsible for considerable losses in farmed and wild fish populations in Europe and North America
Applying the PKD-Recombinase polymerase amplification (RPA) assay combined with an lateral flow dipstick (LFD) enhances the surveillance and early detection of T. bryosalmonae in salmonids
35 zooids were collected from Specific pathogen-free (SPF) F. sultana colonies and subjected to DNA extraction to be used as a negative control
Summary
The myxozoan Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), is responsible for considerable losses in farmed and wild fish populations in Europe and North America. T. bryosalmonae was detected in many European countries, and strategy to control the disease in the wild and farmed fish population is yet to be developed. Proliferative kidney disease (PKD), an emerging parasitic disease, is threatening both farmed and wild salmonid populations in North America and Europe [1, 2]. The malacosporean parasite Tetracapsuloides bryosalmonae is the causative agent of PKD in salmonids [3]. Specific monoclonal antibodies were used for the detection of T. bryosalmonae in the infected fish kidney [32,33,34]. PCR has intrinsic disadvantages, such as timeconsuming, expensive instruments, and complicated procedures for the detection of amplified products [44, 45]
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