Guo X, Chen J, Beuchat L R, Brackett R E
Center for Food Safety and Quality Enhancement, Department of Food Science and Technology, The University of Georgia, Griffin, Georgia 30223-1797, USA.
Appl Environ Microbiol. 2000 Dec;66(12):5248-52. doi: 10.1128/AEM.66.12.5248-5252.2000.
Salmonellae have been some of the most frequently reported etiological agents in fresh-produce-associated outbreaks of human infections in recent years. PCR assays using four innovative pairs of primers derived from hilA and sirA, positive regulators of Salmonella invasive genes, were developed to identify Salmonella enterica serotype Montevideo on and in tomatoes. Based on examination of 83 Salmonella strains and 22 non-Salmonella strains, we concluded that a pair of hilA primers detects Salmonella specifically. The detection limits of the PCR assay were 10(1) and 10(0) CFU/ml after enrichment at 37 degrees C for 6 and 9 h, respectively. When the assay was validated by detecting S. enterica serotype Montevideo in and on artificially inoculated tomatoes, 10(2) and 10(1) CFU/g were detected, respectively, after enrichment for 6 h at 37 degrees C. Our results suggest that the hilA-based PCR assay is sensitive and specific, and can be used for rapid detection of Salmonellae in or on fresh produce.
近年来,沙门氏菌一直是新鲜农产品相关人类感染暴发中报告频率最高的病原体之一。我们开发了基于沙门氏菌侵袭基因的正向调节因子hilA和sirA设计的四对创新引物的PCR检测方法,用于鉴定番茄表面及内部的肠炎沙门氏菌蒙得维的亚血清型。通过对83株沙门氏菌菌株和22株非沙门氏菌菌株的检测,我们得出结论,一对hilA引物能特异性地检测出沙门氏菌。PCR检测方法在37℃富集6小时和9小时后的检测限分别为10¹和10⁰CFU/ml。当通过检测人工接种番茄表面及内部的肠炎沙门氏菌蒙得维的亚血清型来验证该检测方法时,在37℃富集6小时后,分别检测到10²和10¹CFU/g。我们的结果表明,基于hilA的PCR检测方法灵敏且特异,可用于快速检测新鲜农产品表面及内部的沙门氏菌。