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用于检测生牛肉和熟牛肉中肠出血性大肠杆菌O157:H7的视觉免疫沉淀测定八小时法(AOAC官方方法996.09的修订版):协作研究

Visual immunoprecipitate assay eight hour method for detection of enterohemorrhagic Escherichia coli O157:H7 in raw and cooked beef (modification of AOAC Official Method 996.09): collaborative study.

作者信息

Feldsine Philip T, Kerr David E, Leung Stephanie C, Lienau Andrew H, Moser Ruth F, Mui Linda A

机构信息

BioControl Systems, Inc., Bellevue, WA 98005, USA.

出版信息

J AOAC Int. 2002 Sep-Oct;85(5):1029-36.

Abstract

AOAC Official Method 996.09, Visual Immunoprecipitate Assay (VIP) for Escherichia coli O157:H7, was modified to incorporate a new enrichment protocol using BioControl EHEC8 medium for testing raw and cooked beef. Foods were tested by VIP assay and the U.S. Department of Agriculture/Food Safety and Inspection Service (USDA/FSIS) enrichment procedure and the FDA Bacteriological Analytical Manual (BAM) isolation and confirmation techniques. A total of 15 collaborators participated. Raw and cooked ground beef were inoculated with E. coli O157:H7 at 2 different levels: a high level, where predominantly positive results were expected, and a low level where fractional recovery was anticipated. Collaborators tested 396 test portions and controls by both methods, for a total of 792 test portions. Of the 396 paired test portions, 75 were positive and 230 were negative by both the VIP and culture methods. Eleven test portions were presumptively positive by VIP and could not be confirmed culturally; 32 were negative by VIP, but confirmed positive by culture; and 65 were negative by the culture method, but confirmed positive by the VIP method. There was no statistical difference between results obtained with the VIP for EHEC 8 h method and the culture method except for cooked beef, where the VIP had significantly higher recovery for one inoculation level.

摘要

AOAC官方方法996.09,即用于检测大肠杆菌O157:H7的视觉免疫沉淀测定法(VIP),已被修改,纳入了一种使用BioControl EHEC8培养基的新富集方案,用于检测生牛肉和熟牛肉。通过VIP测定法、美国农业部/食品安全与检验局(USDA/FSIS)富集程序以及美国食品药品监督管理局《细菌学分析手册》(BAM)的分离和确认技术对食品进行检测。共有15名合作者参与。将大肠杆菌O157:H7以两种不同水平接种到生牛肉末和熟牛肉末中:一种是高水平,预计主要得到阳性结果;另一种是低水平,预期回收率较低。合作者通过这两种方法对396个测试部分和对照进行了检测,总共792个测试部分。在396对测试部分中,VIP和培养法均为阳性的有75个,均为阴性的有230个。11个测试部分通过VIP初步判定为阳性,但培养法无法确认;32个通过VIP为阴性,但培养法确认为阳性;65个通过培养法为阴性,但通过VIP法确认为阳性。除了熟牛肉外,EHEC 8小时VIP法和培养法的检测结果之间没有统计学差异,在熟牛肉的一个接种水平上,VIP的回收率显著更高。

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