Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/51267
標題: 市售食品及乳房炎病原菌檢測用生物晶片之評估改進及其應用
Evaluation and Improvement of CommercialMicroarray Kits for the Detection of Food and Mastitis Pathogenic Bacteria in Food Samples
作者: Chen, Tzu-Chi
陳姿綺
關鍵字: 食品病原菌
Food and Mastitis Pathogenic Bacteria
乳房炎病原菌
生物晶片套組
Microarray Kits
出版社: 食品暨應用生物科技學系
摘要: 腸炎弧菌、金黃色葡萄球菌、仙人掌桿菌群、沙門氏菌、病原性大腸桿菌等為造成台灣地區食品中毒之常見病原菌,而牛乳房炎則是乳牛場中最常見的疾病,可造成酪農產業嚴重的經濟損失。傳統檢測法先利用選擇性與鑑別性培養基挑出可疑菌落,再進行生化試驗以及血清型鑑定等步驟,費力且費時,故發展一套快速檢測方法有其重要性。生物晶片即為目前先進的技術之一,本研究針對國內業界既有之食品病原菌及牛乳房炎病原菌之生物晶片檢測套組進行檢測正確性、改良與簡化產品降低成本進行評估;並提出其缺點及給予適當之建議。 本論文實驗結果,利用DR. Food-10TM Kit進行純菌系統評估食品生物晶片檢測套組之特異性之試驗,受試結果顯示以 Salmonella spp.、 Escherichia coli、Shigella spp.、Campylobacter spp.、 Vibrio spp.等測試菌株,具有良好之特異性。以四種菌株包括Salmonella Typhimurium、E. coli、Staphylococcus aureus、Bacillus cereus以等菌量混菌,進行DR. Food-10TM Kit試驗,初步試驗結果並無cross hybridization產生。以簡易型DR. FoodTM Kit進行食品系統中之靈敏度試驗,針對沙門氏菌而言,晶片之檢測靈敏度不足,晶片探針之製作條件與樣品預培養條件需加以改良。以DR. FoodTM Kit,進行食品生物晶片簡易型檢測套組評估,檢測菌株包括Salmonella spp.、 E. coli / Shigella spp.、 Listeria monocytogenes,增殖後檢測靈敏度靈可達原始菌數100。乳房炎晶片檢測部分,單獨一株菌用很多雜交點決定,常有部分雜交點未出現之現象,應用於牛乳檢測時,亦有檢測靈敏度不足之問題。因此對於此食品生物晶片檢測套組擬向公司建議,針對菌株特異性差之探針,進行改良,並建議將具有良好特異性之探針發展為簡易型、價廉之細菌檢測之晶片。並請廠商改善生產之晶片可靠性不足,以及儀器穩定性改進等問題。
Abstract Vibrio parahaemolyticus, Staphylococcus aureus, Bacillus cereus group, Salmonella spp., pathogenic Escherichia coli are common pathogens which may cause food poisoning cases. On the other hand, mastitis is considered to be a common disease in dairy herds. Conventional methods for the detection of these bacteria spp. need the use of selection and differentiation medium followed by biochemical and serological identification steps. Such process is time consuming and laborious. Thus, development of rapid methods for the detection of these bacteria species is important. Biochip which allows the simultaneous detection of multiple bacteria spp. has been recently developed and commercialized for this purpose. In this study, we will evaluate the specificity and reliability of the bacteria specific biochips developed domestically and furthermore, improve and modify the biochip if necessary. Results show that the Dr. Food-10TM kit biochip could allow the detection of Salmonella spp.、 Escherichia coli、Shigella spp.、 Campylobacter spp. and Vibrio spp. etc. As this Dr. Food-10TM chip was used for the detection of Salmonella Typhimurium, E. coli, Staphylococcus aureus, Bacillus cereus mixed in equal cell numbers, no cross reaction results. As the detection limit of an economic and simplified chip which allow the detection of Salmonella spp.、 E. coli / Shigella spp.、 Listeria monocytogenes was evaluated, we found that for Salmonella detection, the chip was not sensitive enough. We suggested that either the process for chip preparation or the conditions for culture of target cells in samples should be improved. However, if an enrichment step for target cells was performed prior to the detection, the detection limit for these bacteria became 100. As for the detection of mastitis bacteria, identification of one bacteria spp. usually need several hybridization dots. Under such conditions, some hybridization signal was ambiguous, thus, the detection limit need to be improved, In conclusion, our suggestions to the biochip company are as follows, (1) Improvement of the specificity of the probes for some bacteria species, (2) Development of economic and simplified type of biochips, (3) The process for biochip manufacture and the reliability of instrument used for biochip detection may need to be improved too.
URI: http://hdl.handle.net/11455/51267
Appears in Collections:食品暨應用生物科技學系

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