應用具特異性聚合酶鏈反應（PCR）檢測食品中志賀氏桿菌（Shigella）   全文下載

A Rapid and Specific PCR Method for the Detection of Shigella spp. in Spiked Samples

本研究目的主要發展快速且具特異性之PCR方法，檢測食品中志賀氏桿菌（Shigella spp.）及腸入侵性大腸桿菌（enteroinvasive Escherichia coli, EIEC），PCR使用二組引子VirF2-VirF3及VirF3-VirF4應用於志賀氏桿菌及腸入侵性大腸桿菌檢測，分別得到443 bp 及331 bp大小之PCR產物。食品中接入志賀氏桿菌（101 cells/g）經培養後，經PCR引子檢測可得到正反應。進一步以PCR檢測100種天然樣品中志賀氏桿菌及腸入侵性大腸桿菌。由結果顯示，PCR方法與傳統方法（BAM）皆無檢出志賀氏桿菌及腸入侵性大腸桿菌。"

The purpose of this study was to develop a rapid and specific PCR system to detect Shigella spp in samples. A set of primers specific for the virulence gene (VirF) of virulent Shigella spp. and enteroinvasive Escherichia coli (EIEC) produced specific amplicons of expected sizes of 443 bp and 331 bp by using VirF2-VirF3 and VirF3-VirF4 primer sets, respectively. These primers were then used for the detection of food with 101 cells/g inoculation of Shigella spp., followed by Shigella broth incubation. The presence of this pathogen in artificial contaminated foods was detected. Finally, we used this method for the detection of 100 samples, and found that none of Shigella spp. and EIEC bacterial strains was detected in all tested samples by PCR method and the Bacteriological Analytical Manu (BAM) method. The results indicate that the described PCR method has the advantage of a rapid, sensitivity and specificity for use in natural samples.