Abstract

Aims: FtsZ is a highly conserved protein that is found in most of the major groups of bacteria. Considering the importance of the FtsZ gene in bacteria, in this study, Acinetobacter baumannii, Escherichia coli, and Staphylococcus aureus pathogens were detected by a new degenerate primer using PCR method with the targeting of FtsZ gene. Methods: Of each bacterium 30 clinical samples were collected from clinical diagnostic laboratories. For the specificity, The FtsZ gene was amplified using uniplex and triplex PCR with a degenerate primer as forward and three specific reverse primers for A. baumannii, E. coli, and S. aureus samples and some other species as negative control samples. The sensitivity of the primers was determined by PCR using a serial dilution of the genomic DNA template (100 ng to 1 pg). Results: The results showed that the PCR assay is positive for all of the A. baumannii, E. coli, and S. aureus samples (100%) while the PCR amplification was negative for other species. Conclusions: According to the results, the developed PCR assay using FtsZ gene is a reliable, fast, easy to perform and high specific and sensitive diagnostic method for early detection of the A. baumannii, E. coli, and S. aureus.

中文翻译：

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基于简并引物的PCR，以FtsZ基因为靶标，可快速，准确地检测传染病原体，鲍曼不动杆菌和金黄色葡萄球菌

摘要

目的：FtsZ是一种高度保守的蛋白质，可在大多数主要细菌种类中找到。考虑到FtsZ基因在细菌中的重要性，在这项研究中，使用针对FtsZ基因的PCR方法，通过新型简并引物检测了鲍曼不动杆菌，大肠杆菌和金黄色葡萄球菌病原体。方法：从临床诊断实验室收集每种细菌的30份临床样品。对于特异性，该FtsZ的基因用扩增单重和三重PCR用简并引物作为正向和三特异性反向引物鲍曼不动杆菌，大肠杆菌和金黄色葡萄球菌样品以及其他一些物种作为阴性对照样品。使用基因组DNA模板的系列稀释液（100 ng至1 pg）通过PCR测定引物的敏感性。结果：结果显示，所有鲍曼不动杆菌，大肠杆菌和金黄色葡萄球菌样品（100％）的PCR检测均为阳性，而其他物种的PCR扩增均为阴性。结论：根据这一结果，使用开发PCR测定FtsZ的基因是一种可靠，快速，容易进行和高比和用于早期检测的灵敏的诊断方法鲍曼不动杆菌，大肠杆菌和金黄色葡萄球菌。