[1]边 宇,钱宏伟,孟庆峰,等.框镜鲤致病性维氏气单胞菌双重PCR检测方法的建立[J].中国预防兽医学报,2013,(04):304-307.[doi:doi: 10.3969/j.issn.1008-0589.2013.04.12]
 BIAN Yu,QIAN Hong-wei,MENG Qing-feng,et al.Development of duplex PCR for detection of Aeromonas veronii[J].Chinese journal of preventive veterinary medicine,2013,(04):304-307.[doi:doi: 10.3969/j.issn.1008-0589.2013.04.12]
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框镜鲤致病性维氏气单胞菌双重PCR检测方法的建立
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2013年04期
页码:
304-307
栏目:
诊断和检测技术
出版日期:
2013-04-15

文章信息/Info

Title:
Development of duplex PCR for detection of Aeromonas veronii
文章编号:
1008-0589(2013)04-0304-04
作者:
边 宇1钱宏伟2孟庆峰3贺德聪4比尔来西肯·赛都力15单晓枫1*康元环1王伟利3钱爱东1*
1. 吉林农业大学 动物科学技术学院,吉林 长春 130118;2. 吉林电力医院,吉林 长春 130022;3. 吉林出入境检验检疫局,
吉林 长春 130062;4. 吉林省通化市畜牧总站,吉林 通化 134000;5. 伊犁职业技术学院,新疆 伊犁 835000)
Author(s):
BIAN Yu1 QIAN Hong-wei2 MENG Qing-feng3 HE De-cong4 Bierlaixiken-Saiduli15SHAN Xiao-feng1* KANG Yuan-huan1 WANG Wei-li3 QIAN Ai-dong1*
(1. College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China; 2. The Electric Power Hosptial of Jilin Province, Changchun 130022, China; 3. Jilin Entry-Exit Inspection and Quarantine Bureau, Changchun 130062, China;
4. Tonghua Animal Husbandry Stationary, Tonghua 134000, China; 5. Yili Polytechnic College, Yili 835000, China)
关键词:
维氏气单胞菌气溶素基因16S rRNA双重PCR
Keywords:
Aeromonas veronii Aerolysin gene (Aer) 16S rRNA duplex PCR
分类号:
S852.61
DOI:
doi: 10.3969/j.issn.1008-0589.2013.04.12
文献标志码:
A
摘要:
为建立快速检测框镜鲤致病性维氏气单胞菌(A.veronii)双重PCR方法,本研究以A.veronii CY0806株和国际标准株DNA为模板,分别以16S rRNA和Aer基因特异性引物进行PCR扩增,分别获得大小约880 bp和430 bp的DNA片段。通过序列比对分析,16S rRNA基因片段、Aer片段序列与GenBank中登录的A.veronii ATCC35624株的的同源性均为99 %。进一步试验显示该方法的敏感性较高,达到1.58×10-3 ng/μL,特异性较强,只有A.veronii标准株及分离株结果呈阳性;人工模拟污染样本试验显示:该方法的检出率达到了86.7 %,高于细菌分离培养的70 %检出率。双重PCR检测方法的建立,为框镜鲤致病性A.veronii的检测提供新的方法。
Abstract:
To establish the duplex PCR method for detection of Aeromonas Veronii in Cyprinus carpio, two DNA fragments about 880 bp and 430 bp were amplified with the specific primers of 16S rRNA gene and Aer gene from the chromosome DNA of A.veronii CY0806 strain and reference strain. Alignment analysis indicated that the 2 DNA fragments shared 99% homology in sequence with the gene of A.veronii ATCC35624. The further experiments indicated that the duplex PCR method was specific for A.veronii with a limit detection of 1.58×10-3 ng bacteria DNA. In addition, tested on 30 artificial infected fish samples showed that the detection rate of the duplex PCR method reached 86.7% which was higher than that of 70% by bacterial isolation method. The establishied duplex PCR provided a sensitive method for the detection of A.veronii infection in Cyprinus carpio.

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备注/Memo

备注/Memo:
收稿日期:2012-04-23
基金项目:国家支撑计划项目(2010BAD04B01);吉林省科技厅项目(20080218);吉林农业大学青年启动基金(200904)
作者简介:边 宇(1988-),女,辽宁本溪人,硕士研究生,主要从事动物微生物学研究.
*通信作者:E-mail:qianaidong0115@163.comsxf1997@163.com
更新日期/Last Update: 2013-04-11