1 Department of Food Science and Nutrition, University of Minnesota, St. Paul, Minnesota 55108.
2 Paradigm Diagnostics, Inc., 800 Transfer Road, Suite 12, St. Paul, Minnesota 55114 (ORCID: http://orcid.org/0000-0002-1740-6459 ).
J Food Prot. 2018 Aug;81(8):1252-1257. doi: 10.4315/0362-028X.JFP-17-520.
Microbiological analysis of ground beef for contamination by both Salmonella and Shiga toxin-producing Escherichia coli (STEC) is performed by the U.S. Department of Agriculture, Food Safety Inspection Service (FSIS), as part of its Performance Standards Verification Testing program. FSIS has established a zero tolerance for STEC serotype O157:H7 and serogroups O26, O45, O103, O111, O121, and O145 because they are regarded as adulterants. The detection and isolation of these specific serogroups presents a technical challenge necessitating time-consuming and costly laboratory procedures that often exceed the technical capabilities of many small internal and reference laboratories. We describe here a method using a novel STEC and Salmonella selective (SSS) broth that allows for simultaneous selective enrichment of STEC and Salmonella sp., providing isolation and detection from the same broth. The method only involves direct plating from beef enrichments to detect suspect isolates that can be easily confirmed by using immunoassays or PCR, rendering the isolation simpler and less costly than the current described methods. In a side-by-side comparison with modified tryptic soy broth (mTSB), the use of SSS broth resulted in primarily isolating STEC and Salmonella sp., while substantially suppressing the growth of other gram-negative Enterobacteriacae by 90%. Significantly more (χ < 3.84) samples containing E. coli O157:H7 and STEC O26, O111, O121, and O145 and a nondifferent (χ > 3.84) number of samples containing STEC O103 and O45 were identified when enriching in SSS broth. Coenrichment using six different Salmonella serovars showed numerically greater but not significant (χ < 3.84) positive samples by using SSS broth compared with mTSB for a majority of serotypes.
美国农业部食品安全检验局(FSIS)对碎牛肉进行微生物分析,以检测沙门氏菌和产志贺毒素大肠杆菌(STEC)的污染情况,这是其绩效标准验证测试计划的一部分。FSIS 对 STEC 血清型 O157:H7 和血清群 O26、O45、O103、O111、O121 和 O145 零容忍,因为它们被视为掺杂物。检测和分离这些特定的血清群带来了技术挑战,需要耗时且昂贵的实验室程序,这些程序通常超出了许多小型内部和参考实验室的技术能力。我们在这里描述了一种使用新型 STEC 和沙门氏菌选择性(SSS)肉汤的方法,该方法允许同时选择性地富集 STEC 和沙门氏菌,从同一肉汤中进行分离和检测。该方法仅涉及从牛肉增菌物中直接平板接种以检测可疑分离株,这些分离株可以通过免疫测定或 PCR 轻松确认,从而使分离比当前描述的方法更简单且成本更低。与改良胰蛋白酶大豆肉汤(mTSB)进行平行比较时,使用 SSS 肉汤主要分离出 STEC 和沙门氏菌,同时将其他革兰氏阴性肠杆菌的生长抑制了 90%。当在 SSS 肉汤中进行增菌时,显著更多(χ<3.84)的含有 E. coli O157:H7 和 STEC O26、O111、O121 和 O145 的样品和数量相当(χ>3.84)的含有 STEC O103 和 O45 的样品被鉴定出来。使用 SSS 肉汤对六种不同的沙门氏菌血清型进行共增菌时,对于大多数血清型,使用 SSS 肉汤的阳性样品数量明显更大(χ<3.84),但无统计学意义。