Fazzeli Hossein, Arabestani Mohammad Reza, Esfahani Bahram Nasr, Khorvash Farzin, Pourshafie Mohammad Reza, Moghim Sharareh, Safaei Hajieh Ghasemian, Faghri Jamshid, Narimani Tahmine
Department of Microbiology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
J Res Med Sci. 2012 Jul;17(7):671-5.
Sepsis is a systemic inflammatory response associated with high mortality rates in the clinical setting. A multiplex endpoint polymerase chain reaction (PCR) based assay for rapid detection of enterobacteriaceae involved in septicemia, which included Internal Control (IC) and 16S rDNA, is presented here. To develop a panel of primers for DNA fragments of 16S rDNA, enterobacteriaceae, IC, and evaluate analytical sensitivity and specificity of the test.
Primers for amplification of enterobacteriaceae, IC, and16S rDNA were designed, and then PCR was performed. Minimal analytical sensitivity was determined by cloning and colony PCR, and specificity was tested on the basis of their respective standard strains. This study is a cross-sectional Model.
Our results showed the rpoB gene as the most promising target for detection of enterobacteriaceae by PCR amplification. Specificity and sensitivity of endpoint PCR were 100%, 100%, and 100%, and 10, 1, and 100 copies/reaction for enterobacteriaceae, IC, and 16S rDNA, respectively.
The molecular panel presented offers the advantage of an easy, reliable, and cost-effective system when compared to other molecular detection methods. However, further evaluation is needed. Our assay holds promising for more rapid pathogens related in clinical sepsis.
脓毒症是一种在临床环境中与高死亡率相关的全身炎症反应。本文介绍了一种基于多重终点聚合酶链反应（PCR）的检测方法，用于快速检测参与败血症的肠杆菌科细菌，该方法包括内部控制（IC）和16S核糖体DNA（rDNA）。旨在开发一组用于16S rDNA、肠杆菌科细菌、IC的DNA片段的引物，并评估该检测方法的分析灵敏度和特异性。
设计用于扩增肠杆菌科细菌、IC和16S rDNA的引物，然后进行PCR。通过克隆和菌落PCR确定最低分析灵敏度，并根据各自的标准菌株测试特异性。本研究为横断面模型。
我们的结果表明，rpoB基因是通过PCR扩增检测肠杆菌科细菌最有前景的靶点。终点PCR对肠杆菌科细菌、IC和16S rDNA的特异性和灵敏度分别为100%、100%和100%，以及10、1和100拷贝/反应。
与其他分子检测方法相比，本文介绍的分子检测方法具有简便、可靠且经济高效的优点。然而，仍需进一步评估。我们的检测方法在临床脓毒症中快速检测更多相关病原体方面具有前景。
