開發表面共振效應搭配拉曼光譜技術用於動物病原快速偵測

Development of rapid detection of animal pathogens by the surface-enhanced Raman scattering system

洪瑞辰(Ruei-Chen Hung) ； 陳嘉琪(Chia-Chi Chen) ； 林壬威(Jen-Wei Lin) ； 莊曉莉(Hsiao-Li Chuang) ； 邱健昭(Chien-Chao Chiu) ； 黃娉敏(Ping-Min Huang) ； 洪瑛璟(Ying-Ching Hung) ； 楊程堯(Cheng-Yao Yang) ； 林家羽(Chia-Yu Lin) ； 李卓臨(Cho-Lin Li) ； 洪紹文(Shao-Wen Hung)

《嘉大農林學報》 16卷1期 (2019/06) Pp. 61-84

摘要

目前畜牧產業已由傳統飼養方式轉向大規模集約化飼養，抗生素與疫苗使用導致複雜的混合感染、繼發感染及非典型病徵表現，早期診斷和預防疾病對畜牧產業管理變得更加重要。因此，需要發展一套新式病原診斷方式來加強實驗室診斷和傳統診斷。最近，表面增強拉曼散射（Surface-Enhanced Raman Scattering，SERS）搭配拉曼光譜已被證明在實驗室可用於快速偵測與鑑定病原，此技術平台未來具有高度潛力，可發展臨床病原快速偵測與鑑別系統，並可減少傳統診斷的時間。因此，於本研究試圖建立一套SERS搭配拉曼光譜的病原診斷系統，用於細菌偵測和鑑定。首先，建立2種細菌性囓齒類動物常見傳染病病原（金黃色葡萄球菌與綠膿桿菌）和2種豬隻細菌（豬肺炎黴漿菌與豬鏈球菌）的拉曼光譜。之後，再將金黃色葡萄球菌與綠膿桿菌分別混合小鼠檢體（血清和糞便）；將豬肺炎黴漿菌與豬鏈球菌分別混合豬檢體（血清、糞便、尿液），並利用已知菌鼠平台進行檢體收集，與搭配SERS進行檢測。結果呈現以4種菌進行SERS檢測，可成功建立各菌之拉曼光譜，並可根據其病原的特徵拉曼波峰，進行細菌的快速鑑定。另一方面，已知菌鼠平台所收集的檢體與空白樣品與目標菌進行混合後之檢體，皆能以SERS成功地鑑定細菌。綜合上述結果，SERS搭配拉曼光譜未來將有機會應用於畜、禽及水產養殖現場，針對現場病原進行快速檢測，即時啟動疾病控制，降低農民的經濟損失。

關鍵字

動物疾病 ； 拉曼光譜 ； 快速偵測 ； 表面增強拉曼散射

並列摘要

This shift away from traditional pig farming to large-scale intensive methods has resulted in the increasing cases of complex, mixed infections, or secondary infections. The early diagnostic and disease prevention become more important for pig farm management. However, antibiotic treatment and atypical diseases symptoms might be interfered clinical diagnosis. In order to perform accurate pathology diagnosis, we need to combine the strengthening laboratory diagnosis and traditional diagnostics. Surface-enhanced Raman Scattering (SERS) spectroscopy has recently been shown to be a potentially powerful whole-organism fingerprinting technique and is used for the rapid identification of bacteria. Biosensors based on SERS hold great promise as a platform for sensitive and rapid detection of bacterial pathogens and decrease time of diagnosis. In this report, we tried to characterize and evaluate a SERS-based diagnostic system for the detection and identification of bacteria in the gnotobiotic mice for rapid detection two common kind of bacteria in rodent (Staphylococcus aureus, Pseudomonas aeruginosa) and two kind of pathogenic bacteria in swine (Mycoplasma hyopneumoniae, Streptococcus suis) which in pooled with swine sera, feces, and urine. Spectra of bacteria recovered from sera, feces, and urine were compared to pure culture bacteria. The results indicated that successful detection, identification, and classification of these bacteria from mice specimens (sera and feces) and swine specimens (sera, feces, and urine) using a SERS-based system were demonstrated. SERS is shown to offer reproducible molecular spectroscopic signatures for analytical applications. The approach may be a new and powerful tool for real-time surveillance of animal bacterial disease pathogens in clinics.

並列關鍵字

Animal diseases ； Raman spectrum ； Rapid detection ； Surface-enhanced Raman scattering