An Efficient Tetraplex Surveillance Tool for Salmonid Pathogens

Frontiers in Microbiology

Tessa Averink¹  Karthiga Kumanan¹  Jane E. Symonds¹  Cara L. Brosnahan²  Kate S. Hutson²  Ulla von Ammon²  Xavier Pochon³ 

[1]Marine Biosecurity, Cawthron Institute, Nelson, New Zealand

[2]

[3]Aquaculture &College of Science and Engineering, James Cook University, Townsville, QLD, Australia

[4]Institute of Marine Science, University of Auckland, Warkworth, New Zealand

关键词: droplet digital PCR;    multiplex assay;    Oncorhynchus tshawytscha;    Chinook salmon;    aquatic animal health;    fish disease;   

DOI  :  10.3389/fmicb.2022.885585

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown