草鱼呼肠孤病毒三重PCR检测方法的建立及其应用

首页 > 过刊浏览>2013年第20卷第2期 >419-426

草鱼呼肠孤病毒三重PCR检测方法的建立及其应用
DOI:
                        
CSTR:
                        
作者:
                        
作者单位:中国水产科学研究院 珠江水产研究所, 广东 广州 510380
作者简介:曾伟伟(1981), 男, 助理研究员, 从事分子病毒研究. E-mail: xww8810303@163.com
通讯作者:
中图分类号:S941
基金项目:现代农业产业技术体系建设专项(CARS-46); 中国水产科学研究院基研科研业务经费项目(2012A05026).

Establishment of multiplex PCR for detection of grass carp reovirus and its application

Author:

Affiliation:

Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380

Fund Project:

摘要

图/表

访问统计

参考文献

相似文献

引证文献

资源附件

文章评论

摘要:

优化反应条件和体系PCR 297 bp DNA型核酸最低量分别约为且某一类型的引物只能检测到同一类型的RT-PCR2011主养区的, 本研究建立的检测方法可以特异性的检测各种类型的草鱼呼肠孤病毒利用该方法进行草鱼出血病初步的流行病学分析表明方法的建立旨在对草鱼出血病的快速诊断和分子流行病学调查提供实用的可操作手段。

Abstract:

Three specific primer pairs were designed based on the conserved sequences of different genotypes of the grass carp reovirus (GCRV). 530 bp, 297 bp, and 196 bp DNA fragments were amplified and through optimizing the reaction conditions and system, a was established to detect the three genotypes simultaneously. Further studies indicate that the multiplex-PCR has good amplification efficiency, specificity, and sensitivity. The detection limit was 260 copies/μL, 190 copies/μL, and 230 copies/μL for The specificity experiment indicated that the primers were strictly genotype-specific. This method provides a significant improvement in the detection, genotyping, and molecular epidemiological The detection results of 86 samples collected from 16 cities in the main grass carp breeding area in China using the established multiplex-PCR method revealed that the positive rates of genotypes I, II, and 9.3%, 45.3%, and 2.3%, respectively. The positive co-infection rate of types I and II was 5.8%, that of types II and was not detected. The multiplex PCR for the three genetypes of GCRV were highly specific for the corresponding virus genotypes, with high sensitivity. Preliminary epidemiological data analysis by multiplex PCR indicates that type II is the most common genotype, and the phenomenon of combined infection of different genotypes are in general population of grass carp.

参考文献

相似文献

引证文献

引用本文

曾伟伟,王庆,王英英,张乐生,刘宝芹,石存斌,吴淑勤.草鱼呼肠孤病毒三重PCR检测方法的建立及其应用[J].中国水产科学,2013,20(2):419-426
ZENG Weiwei, WANG Qing, WANG Yingying, ZHANG Lesheng, LIU Baoqin, SHI Cunbin, WU Shuqin. Establishment of multiplex PCR for detection of grass carp reovirus and its application[J]. Journal of Fishery Sciences of China,2013,20(2):419-426

复制

文章指标

点击次数:
下载次数:
HTML阅读次数:
引用次数:

历史

收稿日期:
最后修改日期:
录用日期:
在线发布日期: 2013-04-08
出版日期:

首页

期刊介绍

编委会

下载专区

期刊浏览

投稿指南

期刊征订

收录情况

联系我们

旧版入口

English

引用本文

相关视频

分享

文章指标

历史

文章二维码