多重PCR与免疫层析法检测肉糜中大肠杆菌O157:H7的敏感性比较。

Acta microbiologica Polonica Pub Date : 2002-01-01
Romuald Gryko, Beata M Sobieszczańska, Peter J Stopa, Michał A Bartoszcze
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引用次数: 0

摘要

本研究比较了多重PCR法和免疫层析法检测肉糜中大肠杆菌O157:H7的灵敏度。接种1 ~ 100个大肠杆菌细胞后,两种方法中的任何一种都可以在肉糜中检测到大肠杆菌O157:H7。富集条件为在37℃或42℃的非选择性培养基(缓冲蛋白胨水)中孵育8小时。用三组引物进行多重PCR分析,并用凝胶电泳分析。Quix免疫色谱分析法是由马里兰州哥伦比亚市新视野诊断公司推出的一种新试剂盒,用于对浓缩肉汤进行免疫分析。两种检测方法的灵敏度相似。结果取决于培养物中特定细菌的浓度,因为没有观察到其他细菌对大肠杆菌O157:H7的比例的影响。这些数据表明,这两种方法或一起使用,当与富集技术相结合时,可以提供一种快速检测这种病原体在肉末样品中的存在的方法。
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Comparison of multiplex PCR, and an immunochromatographic method sensitivity for the detection of Escherichia coli O157:H7 in minced beef.

In this study, the sensitivities of multiplex PCR and an immuno-chromatographic methods to detect Escherichia coli O157:H7 in minced beef were compared. The detection of Escherichia coli O157:H7 in minced beef inoculated with 1-100 cells of this bacterium was possible after enrichment of culture and subsequent analysis by either of the two methods. Enrichment conditions were eight hours of incubation at 37 degrees C or 42 degrees C in a non-selective medium (Buffered Peptone Water). Multiplex PCR analysis was performed using three primer sets with analysis by gel electrophoresis. The Quix immuno-chromatographic assay which is a new kit being marketed by New Horizons Diagnostics, Columbia, MD, was used for immunological analysis of the enriched broths.The sensitivity of both tests was similar. The results depended on the concentration of the specific bacterium in the culture since the influence of the proportion of other bacteria to the E. coli O157:H7 was not observed. The data suggests that either method or used together, when coupled with an enrichment technique, could provide a rapid mean to detect the presence of this pathogen in minced meat samples.

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