Use of PCR to Detect Cytophaga psychrophila from Apparently Healthy Juvenile Ayu and Coho Salmon Eggs
Use of PCR to Detect Cytophaga psychrophila from Apparently Healthy Juvenile Ayu and Coho Salmon Eggs
复制标题
使用 PCR 检测表面健康的香鱼和银鲑鱼幼鱼卵中的嗜冷噬细胞菌
DOI：
10.3147/jsfp.32.169
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发表时间：
1997
期刊：
影响因子：
0.6
通讯作者：
H. Wakabayashi
中科院分区：
文献类型：
--
作者：
S. Izumi;H. Wakabayashi
应用套式聚合酶链式反应(Nest-PCR)技术，检测了马哈鱼幼鱼(Plecoglossus Altivelis)和眼卵(Oncorhynchus Kistch)携带的嗜冷胞噬杆菌。阿育幼鱼在琵琶湖被捕获，并在登陆后不久被冻结。当集装箱到达时，来自美国和日本的Coho鲑鱼卵是在成田机场或孵化场按批次收集的。用CHILEX 100作为提取DNA的介质。利用该方法，利用嗜冷链霉菌PSY1和PSY2的特异性引物对其16S rDNA片段进行了扩增。1995年4月，从24份阿育人肾脏组织中提取的DNA中有5份获得了预期大小的扩增产物。这些鱼之前在泰氏琼脂和IFAT上的培养中被诊断为存在嗜冷梭菌阴性。在11批鲑鱼卵中，有5批经PCR检测出嗜冷链球菌阳性。这五个批次中有四个也被培养和IFAT诊断为阳性。此外，用该方法检测肾标本中嗜冷链球菌的灵敏度为1.5CFU/管。
The nested PCR was used to detect Cytophaga psychrophila carried by juvenile ayu (Plecoglossus altivelis) and eyed eggs of coho salmon (Oncorhynchus kistch). Ayu juveniles were caught in Lake Biwa and frozen shortly after landed. The coho salmon eggs from the USA and Japan were collected on a lot-by-lot basis at Narita airport or hatcheries when the containers arrived. Chelex 100 was used as a medium for extraction of DNA from these samples. Utilizing the PCR, a specific primer pair of PSY1 and PSY2 for C. psychrophila was used to amplify its 16S rDNA segments. An amplification product of the expected size was obtained from 5 of 24 DNA extractions from the kidney of ayu samples in April 1995. These fish had been previously diagnosed as negative for the presence of C. psychrophila by cultivation on TYE agar and IFAT. Five of 11 lots of coho salmon eggs were found to be positive for C. psychrophila by PCR. Four of these five lots had also been diagnosed as positive by cultivation and IFAT. In addition, the sensitivity of the PCR amplification for detecting C. psychrophila in kidney samples of ayu was calculated to be 1.5 CFU per PCR reaction tube.