集美大学学报（自然科学版）2026，Vol.31Issue(3)：264-273,10.DOI:10.19715/j.jmuzr.2026.03.02
一种大黄鱼盾纤毛虫病PCR检测方法的建立与应用
Development and Application of PCR Detection Technology for Large Yellow Croaker Scuticociliatosis
摘要
Abstract
In order to prevent outbreaks of scuticociliatosis of juvenile large yellow croaker(Larimichthys crocea)and ensure that parasite-free juveniles are stocked in sea cages,species-specific primers targeting the 18S rRNA gene sequences of Miamiensis avidus and Uronema marinum were designed to establish a multiplex PCR detection method for scuticociliatosis.The method was comprehensively evaluated through primer specific-ity verification,sensitivity testing,and application to naturally infected samples.The results demonstrated that the method could specifically amplify DNA from M.avidus and U.marinum,without cross-reactivity with non-target pathogens such as Trypanosoma,Pseudomonas plecoglossicida,and Iridovirus,nor with DNA from healthy L.crocea tissues,indicating good specificity.Sensitivity tests revealed that the minimum detectable concentrations for M.avidus and U.marinum were 4.82×103 copies/μL(2.4 ng/L)and 1.67×103 copies/μL(1.3 ng/L),re-spectively,demonstrating high sensitivity.Ten naturally infected croaker samples were tested using the PCR meth-od,and six positive samples were detected in total,toutperforming traditional microscopy which detected only five,hus further confirming its diagnostic accuracy and sensitivity in field samples.关键词
大黄鱼/盾纤毛虫/PCR检测/小亚基核糖体RNAKey words
Larimichthys crocea/scuticociliatosis/PCR test/18S rRNA分类
农业科技引用本文复制引用
金健,程志强,王志勇,张东玲..一种大黄鱼盾纤毛虫病PCR检测方法的建立与应用[J].集美大学学报（自然科学版）,2026,31(3):264-273,10.基金项目
国家自然科学基金项目(32172964) （32172964）
福建省自然科学基金项目(2022J02042) （2022J02042）
厦门市科技补助项目(2024CXY0320) （2024CXY0320）
