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1.58×10-3ng/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。
To establish the duplex PCR method for detection of Aeromonas Veronff in Cyprinus carpio, two DNA fragments about 880 bp and 430 bp were amplified with the specific primers of 16S rRNA gene and Aergene 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 1.58x103 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 Cypnnus carpio.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2013年第4期304-307,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
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)
