青蟹双顺反子病毒-1原位杂交检测方法的建立与应用

根据青蟹双顺反子病毒-1 （mud crab dicistrovirus-1,MCDV-1）基因的保守序列设计1对引物,从感染MCDV-1的青蟹鳃组织中提取RNA为模板进行RT-PCR扩增,得到1 314 bp的cDNA片段.纯化后的PCR产物用地高辛（DIG）标记制备核酸探针,此探针与青蟹呼肠孤病毒（MCRV）、鲍肌肉萎缩症病毒（AbSV）、类疱疹病毒（AbHV）、白斑综合症病毒（WSSV）、虎纹蛙病毒（TFV）、锦鲤疱疹病毒（KHV）等水生动物常见病毒均无交叉反应,可以特异性的检测出MCDV-1,检出MCDV-1的灵敏度为8.58＆#215;107 copies/μL.采用原位杂交检测方法...

Full description

Saved in:
Bibliographic Details
Published in广东农业科学 Vol. 42; no. 11; pp. 130 - 134
Main Author 崔亚婷 倪军 马红玲 许海东 冯娟 申亚阳 郭志勋
Format Journal Article
LanguageChinese
Published 上海海洋大学水产与生命学院,上海201306%广东省水生动物疫病预防控制中心,广东广州,510222%中国水产科学研究院南海水产研究所/农业部水产品加工重点实验室,广东广州,510300 2015
中国水产科学研究院南海水产研究所/农业部水产品加工重点实验室,广东广州 510300
Subjects
原位杂交
地高辛核酸探针
拟穴青蟹
青蟹双顺反子病毒-1
青蟹双顺反子病毒-1
原位杂交
拟穴青蟹
地高辛核酸探针
Online AccessGet full text
ISSN1004-874X

Cover

More Information
Summary:根据青蟹双顺反子病毒-1 （mud crab dicistrovirus-1,MCDV-1）基因的保守序列设计1对引物,从感染MCDV-1的青蟹鳃组织中提取RNA为模板进行RT-PCR扩增,得到1 314 bp的cDNA片段.纯化后的PCR产物用地高辛（DIG）标记制备核酸探针,此探针与青蟹呼肠孤病毒（MCRV）、鲍肌肉萎缩症病毒（AbSV）、类疱疹病毒（AbHV）、白斑综合症病毒（WSSV）、虎纹蛙病毒（TFV）、锦鲤疱疹病毒（KHV）等水生动物常见病毒均无交叉反应,可以特异性的检测出MCDV-1,检出MCDV-1的灵敏度为8.58＆#215;107 copies/μL.采用原位杂交检测方法检测拟穴青蟹组织中MCDV-1的组织分布,结果表明,拟穴青蟹中MCDV-1 RNA的阳性信号主要分布于肝胰腺、神经节、肠道和食管.
Bibliography:44-1267/S
CUI Ya-ting, NI Jun, MA Hong-ling, XU Hai-dong, FENG Juan, SHEN Ya-yang, GUO Zhi-xun （ 1. Key Laboratory of Aquatic Product Processing, Ministry of Agriculture/South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China; 2. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 210306, China; Guangdong Provincial Aquatic Animal Epidemic Disease Prevention and Control Center, Guangzhou 510222, China）
Scylla paramamosain; mud crab dicistrovirus-1; dig-labeled probe; in situ hybridization
Mud crab dicistrovirus-1 was isolated from massive deaths of mud crab and had a strong pathogenicity. Based on the MCDV-1 genome sequences, we designed a pair of primers and obtained a 1 314 bp fragment of MCDV-1 gene by RT-PCR. Consequently, probe was prepared using DIG-labeling. The sensitivity and specificity of probe were analyzed by dot blot hybridization method. Results showed that this probe could detect a minimum of 8.58 × 10^7 copies/μL of MCDV-1
ISSN:1004-874X