Trkov Marija, Avgustin Gorazd
Zootechnical Department, Biotechnical Faculty, University of Ljubljana, Domzale, Slovenia.
Int J Food Microbiol. 2003 Jan 15;80(1):67-75. doi: 10.1016/s0168-1605(02)00138-1.
A molecular method for the detection of Salmonella enterica strains based on 16S rRNA sequence analysis was developed by a modification of the previously described PCR primer 16SFI [J. Appl. Bacteriol. 80 (1996) 659], which was combined with a newly developed primer annealing at the position 66-82. Only approximately two thirds of now determined Salmonella 16S rRNA sequences contained a region identical to the 16SFI primer sequence and the reverse primer 16SIII was also not specific. Combined, these two primers have been claimed to allow the specific detection of all Salmonella; however, in this study, they did not recognize S. bongori and 3 out of 78 tested S. enterica strains. They also identified some of the tested Enterobacter cloacae strains as Salmonella. On the contrary, the new primer pair, MINf and MINr, made it possible to recognize correctly all of the 78 tested S. enterica strains, representing 31 different Salmonella serovars. None of the 23 non-Salmonella strains from the related gamma-proteobacterial genera was incorrectly recognized as belonging to S. enterica.
通过对先前描述的PCR引物16SFI[《应用细菌学杂志》80(1996)659]进行修饰，开发了一种基于16S rRNA序列分析检测肠炎沙门氏菌菌株的分子方法，该引物与新开发的在66-82位退火的引物相结合。目前确定的沙门氏菌16S rRNA序列中只有大约三分之二包含与16SFI引物序列相同的区域，并且反向引物16SIII也不具有特异性。据称，这两种引物结合使用可以特异性检测所有沙门氏菌；然而，在本研究中，它们无法识别邦戈尔沙门氏菌以及78株受试肠炎沙门氏菌菌株中的3株。它们还将一些受试阴沟肠杆菌菌株鉴定为沙门氏菌。相反，新的引物对MINf和MINr能够正确识别所有78株受试肠炎沙门氏菌菌株，这些菌株代表31种不同的沙门氏菌血清型。来自相关γ-变形菌属的23株非沙门氏菌菌株中没有一株被错误地识别为属于肠炎沙门氏菌。
