[1]崔晓莹,李泽宇,李完波,等.一种定量检测大黄鱼感染变形假单胞菌方法的建立与应用[J].集美大学学报(自然版),2019,24(5):328-334.
 CUI Xiaoying,LI Zeyu,LI Wanbo,et al.Establishment and Application of a Quantitative Method for Detecting of Pseudomonas plecoglossicid Infection in Large Yellow Croaker (Larimichthys crocea)[J].Journal of Jimei University,2019,24(5):328-334.
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一种定量检测大黄鱼感染变形假单胞菌方法的建立与应用()
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《集美大学学报(自然版)》[ISSN:1007-7405/CN:35-1186/N]

卷:
第24卷
期数:
2019年第5期
页码:
328-334
栏目:
水产、食品与生物技术
出版日期:
2019-09-28

文章信息/Info

Title:
Establishment and Application of a Quantitative Method for Detecting of Pseudomonas plecoglossicid Infection in Large Yellow Croaker (Larimichthys crocea)
作者:
崔晓莹12李泽宇12李完波12王志勇12
(1.集美大学水产学院,福建 厦门 361021 ;2.农业部东海海水健康养殖重点实验室,福建 厦门 361021 )
Author(s):
CUI Xiaoying12LI Zeyu 12LI Wanbo12WANG Zhiyong12
(1.Fisheries College,Jimei University,Xiamen 361021,China;2.Key Laboratory of Healthy Mariculture for the East China Sea of Ministry of Agriculture,Xiamen 361021,China)
关键词:
大黄鱼变形假单胞菌gyrB基因荧光定量PCR相对带菌量
Keywords:
Larimichthys croceaPseudomonas plecoglossicidgyrB genereal-time quantitative PCRrelative pathogen load
摘要:
变形假单胞菌(Pseudomonas plecoglossicid)是引起大黄鱼(Larimichthys crocea)内脏白点病的主要病原。基于变形假单胞菌的gyrB基因与大黄鱼的单拷贝基因gdf-8分别设计1对特异性引物,对所设计的变形假单胞菌gyrB基因的引物特异性进行了检验,并采用实时荧光定量PCR(qRT-PCR)技术检测了400尾人工浸泡感染变形假单胞菌5 d后的大黄鱼幼鱼脾脏中的病原菌的相对含量。结果显示,基于gyrB基因所设计的引物对变形假单胞菌检测具有良好的特异性,可以用于对变形假单胞菌的定性和定量检测;400尾人工攻毒大黄鱼幼鱼脾脏中变形假单胞菌的相对含量(相对带菌量,用gyrB基因拷贝数与大黄鱼gdf-8基因拷贝数之比表示)变化在5.04×10-7~0.482之间,不同个体之间的相对带菌量差异很大,说明不同个体的大黄鱼对变形假单胞菌感染的抵抗力有很显著的差异。
Abstract:
The main pathogen causing white -spotted syndrome in internal organs in large yellow croaker(Larimichthys crocea) is Pseudomonas plecoglossicid.Two pairs of specific primers based on the sequences of gyrB of P.plecoglossicid and the single copy gene gdf-8 of large yellow croaker were designed in this study,respectively.The specificity of the designed primers was first checked.The relative pathogen load (RPL) in the spleen of 400 juvenile large yellow croakers after 5 days of artificial challenge was then examined by real-time quantitative PCR(qRT-PCR).The results showed that the primers designed based on the gyrB gene have good specificity for detecting P.plecoglossicid and could be used for qualitative and quantitative detection of P.plecoglossicid.The RPL of the spleen in the 400 juvenile large yellow croakers ranged from 5.04 ×10-7 to 0 .482(the relative pathogen load,represented by the ratio of gyrB gene copy number and gdf-8 gene copy number).The dramatic difference of RPL among different individuals indicated that the resistance of different individuals to P.plecoglossicid infection was significantly different.

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更新日期/Last Update: 2019-11-04