The usefulness of molecular techniques to assess the presence of Aeromonas spp. harboring virulence markers in foods
The usefulness of molecular techniques to assess the presence of Aeromonas spp. harboring virulence markers in foods
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DOI：
10.1016/s0168-1605(03)00105-3
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发表时间：
2004-07-15
影响因子：
5.4
通讯作者：
Jemmi, T
中科院分区：
文献类型：
--
作者：
Kingombe, CIB;Huys, G;Jemmi, T
共有78个生的和123个加工和即食零售食品样本被用来评估运动气单胞菌的存在。与基于使用氨苄青霉素-糊精琼脂（ADA）培养基的常规培养方法相比，使用最近描述的PCR方法，用ADA法检测，65/201（32.3%）份样品显示为推定气单胞菌属。菌落，而PCR方法揭示了气单胞菌属的存在。在51/201（25.4%）的检测样品中携带靶向毒力基因。两种方法处理的样品的污染率和致病性气单胞菌的存在显著低于原始样品。一个多相鉴定方法，包括生物化学和分子生物学技术被应用到34个PCR阳性推定气单胞菌菌株的选择。经过脂肪酸甲酯（FAME）分析和扩增片段长度多态性（AFLP）指纹图谱分析，共有33个分离株（97%）可以在DNA杂交组（HG）水平上进行鉴定。这些分离物中的大多数属于气单胞菌HG 3（50%）和维氏气单胞菌温和生物变种（HG 8/10）（38%）。通过PCR-限制性片段长度多态性（PCR-RFLP）指纹分析和PCR-扩增子序列分析（PCR-ASA）从这些菌株中获得的PCR扩增子的分子表征允许将所有菌株分类为已知的PCR-RFLP和PCR-ASA类型。总之，目前的研究结果表明，基于PCR的毒力标记检测，PCR-RFLP和PCR-ASA的组合使用提供了一个快速，灵敏，特异性系统，以评估气单胞菌的存在和流行。在食物样本中含有毒力标记。(C)2003 Elsevier B. V.保留所有权利。
A total of 78 raw and 123 processed and ready-to-eat retail food samples were used to assess the presence of motile Aeromonas spp. harboring virulence genes (cytotoxic enterotoxin and hemolysin genes) using a recently described PCR method in comparison with the conventional cultivation method based on the use of Ampicillin-Dextrin Agar (ADA) medium. With the ADA-based method, 65/201 (32.3%) samples showed presumptive Aeromonas spp. colonies whereas the PCR method revealed the presence of Aeromonas spp. harboring the targeted virulence genes in 51/201 (25.4%) of the tested samples. The rate of contaminated samples and the presence of pathogenic Aeromonas were significantly lower with both methods for processed than in case of raw samples. A polyphasic identification approach including biochemical and molecular techniques was applied to a selection of 34 PCR-positive presumptive Aeromonas isolates. Following fatty acid methyl ester (FAME) analysis and amplified fragment length polymorphism (AFLP) fingerprinting, a total of 33 isolates (97%) could be identified to the DNA hybridization group (HG) level. The majority of these isolates belonged to the species Aeromonas hydrophila HG3 (50%) and Aeromonas veronii biovar sobria (HG8/10) (38%). Molecular characterization of PCR amplicons obtained from these strains by PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) fingerprinting and PCR-Amplicon Sequence Analysis (PCR-ASA) allowed classification of all strains in a known PCR-RFLP and PCR-ASA type. In conclusion, the current findings demonstrate that the combined use of PCR-based virulence marker detection, PCR-RFLP and PCR-ASA offers a rapid, sensitive, and specific system to assess the presence and prevalence of Aeromonas spp. harboring virulence markers in food samples. (C) 2003 Elsevier B.V. All rights reserved.