- 學位論文
金奈米粒子免疫色層分析法偵測沙門氏菌之臨床試驗
Detection of Salmonella in the clinical samples using gold nanoparticle and immunochromatography
摘要
沙門氏菌在自然界中為一個相當重要的病原菌，大致上可以分成兩種類型，一種是屬於傷寒型的沙門氏菌感染，造成所謂的傷寒症狀；另一種為非傷寒沙門氏菌，感染會造成腸道炎。沙門氏菌的感染引發急性腸胃炎在台灣地區的夏天為常見的傳染性疾病。在這篇研究裡，我們利用奈米金粒子的視覺特性並且純化沙門氏菌的DNA及RNA，設計針對沙門氏菌在16S rDNA序列的專一探針，以免疫層析法的方式來偵測沙門氏菌，並且將這個方法應用在臨床病人檢體的分析上面，可以快速、方便的來做到沙門氏菌的偵測。在這個實驗中，我們總共收集了1278個病人的糞便檢體，並將這些檢體塗盤在Hektoen agar上面進行初步篩檢，其中有159糞便檢體長出黑色的菌落，以這159個糞便檢體進行進一步的研究分析，同時以vitek 2菌種分析做為比對，結果顯示有80個檢體確實為沙門氏菌感染，偵測的靈敏度也達到了100%，其中有75個檢體也確實為非沙門氏菌感染，偵測的特異性為94.93%。我們設計了這個快速、低成本的實驗，希望可以透過這個檢測方法，取代臨床上對糞便檢體耗時的生化測試。
並列摘要
Salmonella spp. are important bacterial pathogens in the worldwide. In this study, we use the visual characteristics of nano-gold particles and purified Salmonella DNA and RNA, the design of specific probes for Salmonella, analysis of the way to detect Salmonella, and the patients in the clinical application of this method analysis of samples above, we hope the study can quickly and easily to achieve the detection of Salmonella. On Hektoen agar, 159 stool samples containing black or crystalloid colonies were identified using the proposed method. Without any equipment, such as microscope, the red band corresponding Salmonella were observed. After immunochromatography test, 80 of the isolates were correctly identified as Salmonella to achieve 100% sensitivity. 75 samples were correctly identified as non-Salmonella spp., but 4 was incorrectly identified as Salmonella. The specificity is 94.93%. We designed this rapid, convenient, low cost and high sensitivity and specificity experiment, which will help clinical use and promotion.