Ateba Collins Njie, Mbewe Moses
Department of Biological Sciences, North West University, Mafikeng Campus, Private Bag X2046, Mmabatho 2735, South Africa.
Department of Water and Sanitation, University of Limpopo, Turfloop Campus, Private Bag X1106, Sovenga 0727, South Africa.
Int J Mol Sci. 2014 May 30;15(6):9735-47. doi: 10.3390/ijms15069735.
In many developing countries, proper hygiene is not strictly implemented when animals are slaughtered and meat products become contaminated. Contaminated meat may contain Escherichia coli (E. coli) O157:H7 that could cause diseases in humans if these food products are consumed undercooked. In the present study, a total of 94 confirmed E. coli O157:H7 isolates were subjected to the enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction (PCR) typing to generate genetic fingerprints. The ERIC fragments were resolved by electrophoresis on 2% (w/v) agarose gels. The presence, absence and intensity of band data were obtained, exported to Microsoft Excel (Microsoft Office 2003) and used to generate a data matrix. The unweighted pair group method with arithmetic mean (UPGMA) and complete linkage algorithms were used to analyze the percentage of similarity and matrix data. Relationships between the various profiles and/or lanes were expressed as dendrograms. Data from groups of related lanes were compiled and reported on cluster tables. ERIC fragments ranged from one to 15 per isolate, and their sizes varied from 0.25 to 0.771 kb. A large proportion of the isolates produced an ERIC banding pattern with three duplets ranging in sizes from 0.408 to 0.628 kb. Eight major clusters (I-VIII) were identified. Overall, the remarkable similarities (72% to 91%) between the ERIC profiles for the isolate from animal species and their corresponding food products indicated some form of contamination, which may not exclude those at the level of the abattoirs. These results reveal that ERIC PCR analysis can be reliable in comparing the genetic profiles of E. coli O157:H7 from different sources in the North-West Province of South Africa.
在许多发展中国家，动物屠宰时未严格执行适当的卫生措施，导致肉类产品受到污染。受污染的肉类可能含有大肠杆菌O157:H7，如果这些食品未煮熟就被食用，可能会导致人类患病。在本研究中，总共94株确诊的大肠杆菌O157:H7分离株接受了肠杆菌基因间重复一致序列（ERIC）聚合酶链反应（PCR）分型，以生成基因指纹图谱。ERIC片段通过在2%（w/v）琼脂糖凝胶上进行电泳分离。获取条带数据的有无及强度，导出到Microsoft Excel（Microsoft Office 2003）并用于生成数据矩阵。采用非加权组平均法（UPGMA）和完全连锁算法分析相似性百分比和矩阵数据。不同图谱和/或泳道之间的关系以树状图表示。将相关泳道组的数据汇总并报告在聚类表上。每个分离株的ERIC片段数量从1个到15个不等，其大小在0.25至0.771 kb之间变化。很大一部分分离株产生了一种ERIC条带模式，有三个双联体，大小在0.408至0.628 kb之间。鉴定出八个主要聚类（I - VIII）。总体而言，动物源分离株与其相应食品的ERIC图谱之间存在显著相似性（72%至91%），表明存在某种形式的污染，这可能不排除屠宰场层面的污染。这些结果表明，ERIC PCR分析在比较南非西北省不同来源的大肠杆菌O157:H7基因图谱时是可靠的。
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