An Efficient Tetraplex Surveillance Tool for Salmonid Pathogens.

Autor:	von Ammon U; Aquaculture & Marine Biosecurity, Cawthron Institute, Nelson, New Zealand., Averink T; Aquaculture & Marine Biosecurity, Cawthron Institute, Nelson, New Zealand., Kumanan K; Aquaculture & Marine Biosecurity, Cawthron Institute, Nelson, New Zealand.; College of Science and Engineering, James Cook University, Townsville, QLD, Australia., Brosnahan CL; Institute of Marine Science, University of Auckland, Warkworth, New Zealand., Pochon X; Aquaculture & Marine Biosecurity, Cawthron Institute, Nelson, New Zealand.; Animal Health Laboratory, Ministry for Primary Industries, Upper Hutt, New Zealand., Hutson KS; Aquaculture & Marine Biosecurity, Cawthron Institute, Nelson, New Zealand.; College of Science and Engineering, James Cook University, Townsville, QLD, Australia., Symonds JE; Aquaculture & Marine Biosecurity, Cawthron Institute, Nelson, New Zealand.
Jazyk:	angličtina
Zdroj:	Frontiers in microbiology [Front Microbiol] 2022 Apr 21; Vol. 13, pp. 885585. Date of Electronic Publication: 2022 Apr 21 (Print Publication: 2022).
DOI:	10.3389/fmicb.2022.885585
Abstrakt:	Fish disease surveillance methods can be complicated and time consuming, which limits their value for timely intervention strategies on aquaculture farms. Novel molecular-based assays using droplet digital Polymerase Chain Reaction (ddPCR) can produce immediate results and enable high sample throughput with the ability to multiplex several targets using different fluorescent dyes. A ddPCR tetraplex assay was developed for priority salmon diseases for farmers in New Zealand including New Zealand Rickettsia -like organism 1 (NZ-RLO1), NZ-RLO2, Tenacibaculum maritimum , and Yersinia ruckeri . The limit of detection in singleplex and tetraplex assays was reached for most targets at 10 -9  ng/μl with, respectively, NZ-RLO1 = 0.931 and 0.14 copies/μl, NZ-RLO2 = 0.162 and 0.21 copies/μl, T. maritimum = 0.345 and 0.93 copies/μl, while the limit of detection for Y. ruckeri was 10 -8 with 1.0 copies/μl and 0.7 copies/μl. While specificity of primers was demonstrated in previous studies, we detected cross-reactivity of T. maritimum with some strains of Tenacibaculum dicentrarchi and Y. ruckeri with Serratia liquefaciens , respectively. The tetraplex assay was applied as part of a commercial fish disease surveillance program in New Zealand for 1 year to demonstrate the applicability of tetraplex tools for the salmonid aquaculture industry. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2022 von Ammon, Averink, Kumanan, Brosnahan, Pochon, Hutson and Symonds.)
Databáze:	MEDLINE
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