Coburn J T, Lytle F E, Huber D M
Anal Biochem. 1986 Apr;154(1):305-10. doi: 10.1016/0003-2697(86)90529-4.
Laser-excited fluorimetry has been applied to the identification of bacteria and fungus. The instrumental sensitivity and selectivity of the aminopeptidase profiling method has been enhanced by the use of laser excitation in conjunction with improved spectral and temporal background rejection. The linear dynamic range for the aminopeptidase technique has been increased by achieving a reduced lower limit of detection of the fluorescent tag, beta-naphthylamine. Standard aminopeptidase methodology only provides a linear dynamic range of 1.5 orders of magnitude. The laser-based method expanded the range to three orders of magnitude allowing the inherent specificity of aminopeptidase enzymes within the pathogen to be observed. The enhanced linear dynamic range was observed in profiles of Agrobacterium tumefaciens rubi and Phytophthora megasperma var. sojae.
激光激发荧光法已应用于细菌和真菌的鉴定。通过结合激光激发以及改进的光谱和时间背景抑制技术,氨肽酶分析方法的仪器灵敏度和选择性得到了提高。通过降低荧光标记物β-萘胺的检测下限,氨肽酶技术的线性动态范围得以扩大。标准氨肽酶方法的线性动态范围仅为1.5个数量级。基于激光的方法将范围扩大到三个数量级,从而能够观察到病原体中氨肽酶的固有特异性。在根癌土壤杆菌悬钩子变种和大豆疫霉大豆变种的分析中观察到了增强的线性动态范围。