Naravaneni Rambabu, Jamil Kaiser
Department of Genetics, Bhagwan Mahavir Hospital and Research Centre, Masab tank, Hyderabad, Andhra Pradesh, India-500 004.
J Med Microbiol. 2005 Jan;54(Pt 1):51-54. doi: 10.1099/jmm.0.45687-0.
Traditional methods of identification of food-borne pathogens, which cause disease in humans, are time-consuming and laborious, so there is a need for the development of innovative methods for the rapid identification of food-borne pathogens. Recent advances in molecular cloning and recombinant DNA techniques have revolutionized the detection of pathogens in foods. In this study the development of a PCR-based technique for the rapid identification of the food-borne pathogens Salmonella and Escherichia coli was undertaken. Suitable primers were designed based on specific gene fimA of Salmonella and gene afa of pathogenic E. coli for amplification. Agarose gel electrophoresis and subsequent staining with ethidium bromide were used for the identification of PCR products. The size of the amplified product was 120 bp as shown by comparison with marker DNA. These studies have established that fimA and afa primers were specific for detecting Salmonella and pathogenic E. coli, respectively, in the environmental samples. Thus a rapid, sensitive and reliable technique for the detection of Salmonella and pathogenic E. coli was developed.
导致人类患病的食源性病原体的传统鉴定方法既耗时又费力,因此需要开发创新方法来快速鉴定食源性病原体。分子克隆和重组DNA技术的最新进展彻底改变了食品中病原体的检测方法。在本研究中,开展了一种基于聚合酶链反应(PCR)的技术来快速鉴定食源性病原体沙门氏菌和大肠杆菌。基于沙门氏菌的特异性基因fimA和致病性大肠杆菌的afa基因设计了合适的引物用于扩增。采用琼脂糖凝胶电泳及随后用溴化乙锭染色来鉴定PCR产物。与标记DNA比较显示,扩增产物的大小为120碱基对。这些研究证实,fimA和afa引物分别对环境样品中沙门氏菌和致病性大肠杆菌的检测具有特异性。因此,开发出了一种快速、灵敏且可靠的检测沙门氏菌和致病性大肠杆菌的技术。