Razei Ali, Sorouri Rahim, Mousavi Seyed Latif, Nazarian Shahram, Amani Jafar, Aghamollaei Hosien
Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Department of Microbiology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Iran J Basic Med Sci. 2017 Sep;20(9):1050-1055. doi: 10.22038/IJBMS.2017.9275.
and are three toxin producing bacteria over the world, especially in Iran, and it is essential to find a certain, rapid procedure to identify these microorganisms. In this research, these bacteria were simultaneously detected by multiplex PCR technique in foods.
The primary approval of bacterial strains was performed by biochemical tests. PCR primers were designed based on the nucleotide sequences of the gene of . , the gene of and the C gene of . The specificity of Multiplex PCR method was determined using seven food poisoning bacteria including . To confirm the reaction, DNA extraction was performed from 30 food samples (milk), and gene amplification was performed by PCR. The length of amplified fragments was 300 bp, 210 bp and 160 bp for , and genes, respectively.
The detection limits of the PCR method were 5, 4 and 3 pg for , and , respectively. Specifisity test showed that this reaction is spesific to these 3 bacteria.
In this study, we introduced a new multiplex PCR method for simultsnus detection of and . These results can be used for detection of other toxin producing bacteria in food.
[具体细菌名称1]、[具体细菌名称2]和[具体细菌名称3]是世界范围内,尤其是伊朗的三种产毒素细菌,找到一种确定、快速的程序来鉴定这些微生物至关重要。在本研究中,通过多重PCR技术在食品中同时检测这些细菌。
通过生化试验对菌株进行初步鉴定。基于[具体细菌名称1]的[具体基因名称1]基因、[具体细菌名称2]的[具体基因名称2]基因和[具体细菌名称3]的C基因的核苷酸序列设计PCR引物。使用包括[具体细菌名称4]在内的七种食物中毒细菌确定多重PCR方法的特异性。为了确认反应,从30份食品样本(牛奶)中进行DNA提取,并通过PCR进行基因扩增。[具体细菌名称1]、[具体细菌名称2]和[具体细菌名称3]基因扩增片段的长度分别为300 bp、210 bp和160 bp。
PCR方法对[具体细菌名称1]、[具体细菌名称2]和[具体细菌名称3]的检测限分别为5 pg、4 pg和3 pg。特异性试验表明该反应对这三种细菌具有特异性。
在本研究中,我们引入了一种新的多重PCR方法用于同时检测[具体细菌名称1]、[具体细菌名称2]和[具体细菌名称3]。这些结果可用于检测食品中其他产毒素细菌。