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Multilocus Sequence Typing Lacks the Discriminatory Ability of Pulsed-Field Gel Electrophoresis for Typing Salmonella enterica Serovar Typhimurium

机译:多基因座序列分型缺乏脉冲场凝胶电泳区分肠炎沙门氏菌血清型鼠伤寒的鉴别能力。

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摘要

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.
机译:非伤寒沙门氏菌是美国食源性疾病的主要原因。由于肠炎沙门氏菌在食源性疾病中的重要性,因此开发了许多分型方法。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。这项研究的目的是比较在分型肠炎沙门氏菌鼠伤寒沙门氏菌临床分离株中,PFGE与多基因座序列分型(MLST)的鉴别能力。这项研究共使用了85株来自牛的鼠伤寒沙门氏菌临床分离株。通过疾病控制和预防中心的方法对85个分离株进行了使用XbaI的PFGE,并使用BioNumerics软件包对数据进行了分析。在分离物中观察到50个PFGE图谱,并将其分为三个主要簇。对于MLST分析,通过PCR从相同的85个分离物中扩增了manB,pduF,glnA和spaM基因。这四个基因,manB,pduF,glnA和spaM的DNA测序表明,在测试的分离物中没有遗传多样性,核苷酸序列具有100%的同一性。此外,上述基因的DNA序列与GenBank报道的肠炎链球菌鼠伤寒沙门氏菌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鉴别肠炎沙门氏菌血清鼠伤寒的能力。

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