Fakhr Mohamed K, Nolan Lisa K, Logue Catherine M
Department of Veterinary and Microbiological Sciences, North Dakota State University, Fargo, ND 58105, USA.
J Clin Microbiol. 2005 May;43(5):2215-9. doi: 10.1128/JCM.43.5.2215-2219.2005.
Nontyphoidal salmonellae are among the leading causes of food-borne disease in the United States. Because of the importance of Salmonella enterica in food-borne disease, numerous typing methodologies have been developed. Among the several molecular typing methods, pulsed-field gel electrophoresis (PFGE) is currently considered the "gold standard" technique in typing Salmonella. The aim of this study was to compare the discriminatory power of PFGE to multilocus sequence typing (MLST) in typing Salmonella enterica serovar Typhimurium clinical isolates. A total of 85 Salmonella Typhimurium clinical isolates from cattle were used in this study. PFGE using XbaI was performed on the 85 isolates by the Centers for Disease Control and Prevention method, and data were analyzed using the BioNumerics software package. Fifty PFGE profiles were observed among the isolates, and these grouped into three major clusters. For the MLST analysis, the manB, pduF, glnA, and spaM genes were amplified by PCR from the same 85 isolates. DNA sequencing of these four genes, manB, pduF, glnA, and spaM, showed no genetic diversity among the isolates tested, with a 100% identity in nucleotide sequence. Moreover, the DNA sequences of the aforementioned genes showed 100% identity to the sequence reported in GenBank for the S. enterica serovar Typhimurium LT2 strain. Therefore, MLST, using these genes, lacks the discriminatory power of PFGE for typing Salmonella enterica serovar Typhimurium.
非伤寒沙门氏菌是美国食源性疾病的主要病因之一。由于肠炎沙门氏菌在食源性疾病中的重要性,已开发出多种分型方法。在几种分子分型方法中,脉冲场凝胶电泳(PFGE)目前被认为是沙门氏菌分型的“金标准”技术。本研究的目的是比较PFGE与多位点序列分型(MLST)对肠炎沙门氏菌鼠伤寒血清型临床分离株的分型鉴别能力。本研究共使用了85株来自牛的鼠伤寒沙门氏菌临床分离株。按照疾病控制与预防中心的方法,对这85株分离株进行了XbaI酶切的PFGE分析,并使用BioNumerics软件包对数据进行分析。在分离株中观察到50种PFGE图谱,这些图谱分为三个主要簇。对于MLST分析,通过PCR从相同的85株分离株中扩增出manB、pduF、glnA和spaM基因。对这四个基因manB、pduF、glnA和spaM进行DNA测序,结果显示在所测试的分离株中没有遗传多样性,核苷酸序列具有100%的同一性。此外,上述基因的DNA序列与GenBank中报道的肠炎沙门氏菌鼠伤寒血清型LT2菌株的序列具有100%的同一性。因此,使用这些基因的MLST缺乏PFGE对肠炎沙门氏菌鼠伤寒血清型进行分型的鉴别能力。