Identification of virulent Rhodococcus equi by amplification of gene coding for 15- to 17-kilodalton antigens
Identification of virulent Rhodococcus equi by amplification of gene coding for 15- to 17-kilodalton antigens
复制标题
通过扩增 15 至 17 千道尔顿抗原的基因编码来鉴定强毒力马红球菌
DOI：
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发表时间：
1995
影响因子：
9.4
通讯作者：
T. Sekizaki
中科院分区：
文献类型：
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作者：
S. Takai;T. Ikeda;Y. Sasaki;Yukari Watanabe;Toshisuke Ozawa;S. Tsubaki;T. Sekizaki
在用质粒谱和蛋白质谱对马场马红球菌强毒流行情况进行调查时，从马粪中分离到1725株马红球菌，其中66株(3.8%)和土壤中分离到129株(5.9%)不同大小的隐质粒。对含有不同大小的隐质粒的22株分离物进行了质粒图谱分析，并对其蛋白质谱和小鼠致病性进行了检测。在22个分离株中，有7个是马立克氏杆菌强毒株，同时含有强毒和潜伏质粒，表达15-17 kDa的抗原。其余15株无毒力且不表达抗原：6株含有两种不同大小的隐质粒，9株含有不同大小的隐质粒。建立了一种快速鉴定马立克氏杆菌毒力质粒的聚合酶链式反应方法。从马立克氏杆菌15-17 kDa毒力相关抗原的编码基因序列中提取的寡核苷酸引物，从所有携带毒力质粒的受试菌株中扩增出一条564个碱基的产物。在Southern杂交试验中，该扩增产物与毒力质粒DNA杂交。毒力质粒治愈的衍生物和所有只含有隐质粒的受试菌株均为阴性。与常规方法相比，该方法是一种快速、灵敏、特异的检测方法，可用于从环境分离株中鉴定马立克氏杆菌的毒力，并被认为是流行病学研究的有用工具。
During a survey of the prevalence of virulent Rhodococcus equi at horse-breeding farms by plasmid and protein profiles, cryptic plasmids of various sizes were found in 66 (3.8%) of 1,725 isolates from feces of horses and 129 (5.9%) of 2,200 isolates from soil. Twenty-two isolates, which contained cryptic plasmids of different sizes, were found by plasmid profiles, and their protein profiles and mouse pathogenicities were examined. Of the 22 isolates, 7 were virulent R. equi, contained both virulence and cryptic plasmids, and expressed 15- to 17-kDa antigens. The remaining 15 isolates were avirulent and did not express the antigens: 6 strains contained cryptic plasmids of two different sizes and 9 strains contained cryptic plasmids of various sizes. A PCR assay was developed for the rapid identification of virulence plasmids of R. equi. Oligonucleotide primers, derived from the sequence of a gene coding for the 15- to 17-kDa virulence-associated antigens of R. equi, amplified a 564-bp product from all the tested isolates harboring a virulence plasmid. This PCR product hybridized with virulence plasmid DNA in the Southern hybridization assay. Virulence plasmid-cured derivatives and all of the tested isolates harboring cryptic plasmids only were negative. The PCR is a rapid, sensitive, and specific test for the identification of virulent R. equi from environmental isolates compared with standard techniques, such as plasmid and protein profiles and the mouse pathogenicity test, and is considered to be a useful tool for epidemiological studies.