Rapid detection of the fish-pathogenic bacterium Pasteurella piscicida by polymerase chain reaction targetting nucleotide sequences of the species-specific plasmid pZP1.
Rapid detection of the fish-pathogenic bacterium Pasteurella piscicida by polymerase chain reaction targetting nucleotide sequences of the species-specific plasmid pZP1.
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通过针对物种特异性质粒 pZP1 的核苷酸序列的聚合酶链反应快速检测鱼类致病菌杀鱼巴氏杆菌。
DOI：
10.3147/jsfp.32.143
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发表时间：
1997
期刊：
影响因子：
0.6
通讯作者：
I. Hirono
中科院分区：
文献类型：
--
作者：
T. Aoki;D. Ikeda;T. Katagiri;I. Hirono
从鱼类致病性细菌杀鱼巴斯德氏菌中分离到一个种特异性质粒pZP1。从质粒pZP1中克隆了两个PZP1基因片段（PZP1 - 1，964 bp; PZP1 - 4，477 bp）。根据片段PZP1 - 1和PZP1 - 4的核苷酸序列，构建了两个20-mer引物组，PZP1 - 1a和PZP1 - 1b，PZP1 - 4a和PZP1 - 4b。以40株铜绿假单胞菌为模板，扩增出484 bp的DNA片段。使用PZP1 - 1a/1b引物组进行杀鱼性PCR。这些菌株是在日本和美国的不同地方于一年中的不同时间分离的。从上述菌株中均扩增出321 bp的PCR产物。使用PZP1 - 4a/4b引物组，对除ATCC 17911菌株外的杀鱼性菌株进行PCR扩增。对Beneckea proteolytica、Photobacterium damsela、Ph.histaminum、Ph.leiognathi和15株弧菌标准菌株的模板DNA进行PCR扩增，均未得到类似的扩增产物。或4种鱼类致病菌（Aeromonaserebula，A.杀沙门氏菌、迟缓爱德华氏菌和杀seriolicida肠球菌）。用PZP1 - 1a/1b引物对自然感染P.杀鱼的DNA，但不是来自健康的黄尾鱼的DNA。
A species-specific plasmid (pZP1) has been isolated from the fish-pathogenic bacterium Pasteurella piscicida. Two DNA fragments of PZP1 (PZP1-1 with 964 bp and PZP1-4 with 477 bp) were cloned from the plasmidpZP1. Two 20-mer primer sets, PZP1-1a and PZP1-1b, and PZP1-4a and PZP1-4b, were constructed according to the nucleotide sequences of fragments PZP1-1 and PZP1-4. A 484-bp DNA fragment was amplified from template DNA from 40 strains ofP. piscicidaby PCR using the PZP1-1 a/1b primer set. The strains were isolated at different times of the year at different places in Japan and the USA. A 321-bp PCR product was amplified from all of the above strains ofP. piscicidaexcept strain ATCC17911 using the PZP1-4a/4b primer set. No such PCR products were obtained from template DNAs ofBeneckea proteolytica, Photobacterium damsela, Ph. histaminum, Ph. leiognathi, 15standard strains ofVibriospp. or four fish pathogenic bacteria (Aeromonas hydrophila, A. salmonicida, Edwardsiella tarda and Enterococcus seriolicida). The PCR products were amplified with the PZP1-1a/1b primer set from DNA from the kidney of yellowtail (Seriola quinqueradiata) infected naturally withP. piscicida, but not from DNA from healthy yellowtail.
影响因子：
56.9
作者：
SAIKI, RK;SCHARF, S;ARNHEIM, N
通讯作者：
ARNHEIM, N