2023-01-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/674379食品發酵過程中的菌相變化，可能影響其品質與安全性，若發酵條件控制失當可能導致非預期之微生物增生，進而危害人體健康。因此，有必要發展以科學方法分檢測各發酵製程中菌相變化的技術，分病原菌與優勢菌種之消長，以評估並提升發酵製程之食品安全性。 相較於傳統培養分析技術受限於培養基選擇性而無法反映完整菌相組成，近年得益於次世代定序與生物資訊學成熟，發展出總基因體學16S rRNA基因定序之非培養型菌相分析技術，將其結合即時聚合酶鏈式反應定量16S rRNA基因拷貝數以表示絕對菌數，將可反映樣品完整菌相、同步鑑定樣品中所有菌種、分析各菌種絕對數量上之消長，若能應用此技術於分析食品發酵製程之菌相變化與病原菌消長，將有助於提升其安全性。 本計畫選定我國傳統發酵食品台式泡菜為對象，建立以總基因體學16S rRNA基因定序為基礎之菌相分析平台，將其應用於分析食醋之添加於台式泡菜菌相變化，檢測其中病原菌消長以探討醋之添加是否有助提升其安全性。本計畫在食品安全領域具學術新穎性，將此方法學之應用拓展至食品安全管理，並可依分析結果提出符合食品安全要求的發酵條件與製程建議，作為傳統發酵食品生產指引。 Microbial community of a fermentation food is highly correlated with its safety and quality. Inadequate fermentation condition can lead to the growth of undesired microorganisms such as spoilage bacteria and pathogens, which increases the food safety risks. Therefore, to guarantee the safety of fermentation foods, an analytical method for the surveillance of microbial community and pathogen succession during fermentation is urgently essential. Based on the development of next generation sequencing technology and bioinformatic tools, metagenomic-based 16S rRNA gene sequencing is now a powerful technique for microbiome analysis. The combination of microbial community provided by 16S rRNA gene sequencing and the copy number of 16S rRNA gene quantified by real-time polymerase chain reaction (real-time PCR) can reveal the comprehensive succession of various bacteria in the samples. With the application of this method, surveillance for microbiome composition could be achieved, which could further enforce food safety. The aim of this study is to establish a metagenomic-based cultural-independent platform for the surveillance of microbial communities of Taiwanese pickle. The effect of different manufacturing procedures on the microbiome dynamics will be revealed. The analytical platform will be established, and it will be applied to illuminate the impact of vinegar-added fermentation on the pathogen succession and safety of Taiwanese pickle. This study aims to extend the applicability of metagenomic strategy to food safety. The results could provide recommendations for fermentation conditions and manufacturing procedures, which serve as the guideline for the production of traditional fermentation foods.非經培養法;16S核糖體RNA基因;次世代定序;即時聚合酶鏈式反應;微生物定量;culture-independent method; 16S rRNA gene; next generation sequencing; real-time PCR; microorganism quantification