致病性迟钝爱德华氏菌毒力基因的PCR检测  

Detection of the Virulence Genes of Pathogenic Edwardsiella Tarda by PCR Assay

作　　者：江云[1] 李寿崧[2] 王寿昆[1] 郭立新[2] 江树勋[2] 陈文炳[2] 邵碧英[2] 

机构地区：[1]福建农林大学动物科学学院,福州350002 [2]福建出入境检验检疫局,福州350001

出　　处：《中国食品学报》2008年第4期123-129,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：福建省科技厅重点项目(2004Y046)

摘　　要：为了筛选可用于检测致病性迟钝爱德华氏菌的毒力基因,最终建立致病性迟钝爱德华氏菌PCR检测体系,根据致病性迟钝爱德华氏菌(Edwardsiella tarda,E.et w.)PPD130/91分离株的6个毒力基因citC、fimA、gadB、katB、mukF、esrB序列,设计了6对引物,进行PCR扩增、引物灵敏度试验及特异性试验。结果显示:在国内分离的3个致病株中同时出现citC、fimA、gadB、mukF4个毒力基因,而katB和esrB毒力基因未出现。各毒力基因单重PCR灵敏度试验中,gadB基因可检测到的模板量为1pg,其余3个毒力基因可检测到的模板量为100pg。在特异性试验中,11株常见致病菌没有扩增出与目的基因大小相近的片段,引物特异性良好。由此推断,citC、fimA、gadB、mukF4个毒力基因中的任意一个,均可作为检测致病性迟钝爱德华氏菌的标志物,但此结论还有待于更多国内分离株的验证。In order to screen the virulence genes of pathogenic Edwardsiella tarda, which could be used to detect pathogenic E.tarda, and perform the PCR detection system of pathogenic E.tarda finally, six pairs of primers were designed according to the published nucleotide sequence of virulence genes（orfA, citC, fimA, gadB, katB, mukF and esrB） of PPD130/91 isolation strains. PCR assay, the sensitivity assay and the specificity assay were developed. The result indicated that citC, fimA, gadB and mukF genes were detected simultaneity in domestic isolation strains of pathogenic E.tarda, and katB and esrB genes were not. In the sensitivity assay, the sensitivity of gadB reached lpg, and others reached 100 pg by simple PCR. The four pairs of primers were specific in other eleven familiar pathogenic bacterium, not presenting close size bands. Thus, deduce that any of the four virulence genes can be used as marker to detect the pathogenic E.tarda. The conclusion will need confirmation of more domestic isolation strains.

关 键 词：致病性迟钝爱德华氏菌 毒力基因 PCR检测 

分 类 号：TS207.4[轻工技术与工程—食品科学]