An Efficient Tetraplex Surveillance Tool for Salmonid Pathogens

von Ammon, Ulla, Averink, Tessa, Kumanan, Karthiga, Brosnahan, Cara L., Pochon, Xavier, Hutson, Kate S., and Symonds, Jane E. (2022) An Efficient Tetraplex Surveillance Tool for Salmonid Pathogens. Frontiers In Microbiology, 13. 885585.

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Abstract

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.

Item ID: 74533
Item Type: Article (Research - C1)
ISSN: 1664-302X
Keywords: aquatic animal health, Chinook salmon, droplet digital PCR, fish disease, multiplex assay, Oncorhynchus tshawytscha
Copyright Information: © 2022 von Ammon, Averink, Kumanan, Brosnahan, Pochon, Hutson and Symonds. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Date Deposited: 03 Nov 2022 00:49
FoR Codes: 30 AGRICULTURAL, VETERINARY AND FOOD SCIENCES > 3005 Fisheries sciences > 300503 Fish pests and diseases @ 50%
31 BIOLOGICAL SCIENCES > 3107 Microbiology > 310799 Microbiology not elsewhere classified @ 50%
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