中国海洋大学学报（自然科学版）2024，Vol.54Issue(5)：44-51,8.DOI:10.16441/j.cnki.hdxb.20230102
卵形鲳鲹源海豚链球菌检测方法的建立及应用
Establishment and Application of Detection Method of Streptococcus iniae from Trachinotus ovatus
摘要
Abstract
For analyzing the Streptococcus iniae load of tissue simple rapidly,the specific gene SimA of S.iniae was selected as the target in this study,and a real-time fluorescence absolute quantitative method detecting S.iniae was established.The minimal detection value of this method was 1.93×102 copies/μL with excellent discrimination between 1.93×102 and 1.93×109 copies/μL.The coefficient of variance of repeated experiments was less than 1%,thus the method was highly reproducible.This method can specifically detect S.iniae without interference of ten pathogenic bacteria including V.canadensis,V.alginolyticus,V.harveyi,V.parahaemolyticus among others.According to the absolute quantitative method,the bacterial load of S.iniae in the tissues was analyzed after immersion infected T.ovatus.The load of S.iniae displayed an increasing trend with the extension of infection time in liver,spleen,heart,kidney,brain,gill,and intestine of T.ovatus.Meanwhile,the bacterial load each tissue was quite different.At 12 to 24 h after infection,the tissue with the highest bacterial load was brain,and that with the lowest was intestine.At 48 to 72 h after infection,the tissue with the highest bacterial load was kidney,and that with the lowest was gill.According to the analysis of bacterial load each tissue,S.iniae had the strongest tropism to the brain and kidney of T.ovatus,and relatively weak tropism to the spleen,liver,intestine,heart and gills.In summary,the detection method established in this study was applicable for the dynamic monitoring of infected S.iniae on T.ovatus,which provided a reliable basis for the prevention of S.iniae.关键词
海豚链球菌/卵形鲳鲹/绝对定量/检测/组织嗜性Key words
Streptococcus iniae/Trachinotus ovatus/absolute quantitation/detection/tissue tropism分类
农业科技引用本文复制引用
李姿懿,孙敏敏,王义芬,王茁宇,牛晶晶,宋建强,贺艳,齐洁..卵形鲳鲹源海豚链球菌检测方法的建立及应用[J].中国海洋大学学报（自然科学版）,2024,54(5):44-51,8.基金项目
海南省重点研究发展项目(ZDYF2022XDNY137)资助 Supported by the Key Research and Development Project of Hainan Province(ZDYF2022XDNY137) （ZDYF2022XDNY137）
