食品中弓形菌16S rRNA特异性扩增检测方法的建立  

Development of a specific amplification method of Arcobacter 16S rRNA gene in foods

作　　者：游淑珠[1] 王小玉[1] 胡松楠[1] 唐食明[1] 成晓维[1] 邝筱珊[1] 冯家望[1] 

机构地区：[1]珠海出入境检验检疫局技术中心,珠海519015

出　　处：《食品科技》2011年第10期293-296,共4页Food Science and Technology

基　　金：2011年度国家质检总局科研计划项目(2011IK256)

摘　　要：针对弓形菌16SrRNA基因合成1对引物,通过对聚合酶链式反应(PCR)扩增条件的优化,建立了检测弓形菌的PCR方法。3株弓形菌标准菌株PCR产物测序结果与NCBI上公布的弓形菌16S rRNA基因序列进行比对,比对结果表明3株弓形菌测序结果与NCBI上公布的弓形菌16S rRNA基因序列同源性均在99%以上。3株弓形菌标准菌株均特异性地扩增出了长度为1202bp的片段,其他19株不同种类的菌株均无扩增产物出现。55份食品样品用Johnson-Murano肉汤增菌后用此法进行检测,其中6份样品为弓形菌阳性,阳性率为10.9%。上述实验结果表明,方法特异性强、操作简便,节省了检测时间,可用于食品中弓形菌的快速检测。One pair of primers were used to amplify 16S rRNA sequence of Arcobacter,and a PCR method for detection of Arcobacter was developed with optimization of PCR amplification conditions.PCR products of 3 different Arcobacter type strains were sequenced and compared with 16S rRNA genes of Arcobacter published on NCBI.It＇s proved that the homology between the PCR products and 16S rRNA genes of Arcobacter published were all above 99%.3 different type strains of Arcobacter produced a 1202 bp amplified band,while all of the other 19 different bacteria didn＇t produced any band.55 food samples were detected whether Arcobacter present or not by the PCR method following enrichment in Johnson-Murano broth,and 6 samples were detected positive for this microorganism,namely with the positive ratio of 10.9%.The result suggested that the method developed was specific,easy to operate and timesaving,and could be used for rapid detection of Arcobacter in foods.

关 键 词：弓形菌 16S RRNA 聚合酶链式反应 

分 类 号：Q93-33[生物学—微生物学]