光学 精密工程, 2010, 18 (4): 771, 网络出版: 2010-08-31
生物发光法细菌快速检测仪的研制及应用
Development and application of rapid detecting instrument for bacteria based on bioluminescence
细胞检测仪 生物发光 快速检测 三磷酸腺苷(ATP) detecting instrument of bacterium bioluminescence rapid detection Adenosine Triphosphate(ATP)
摘要
基于ATP生物发光法,研制了一种细菌快速检测仪。该检测仪内置高精度A/D转换器,以小型光电倍增管进行光电转换,结合实验室自制的生物传感器,集光路、电路及软件设计于一体,组成了完整的细菌快速检测系统。其用于大肠杆菌标准品溶液检测时,检测结果与培养计数法结果的相关系数(R)达到0.976 0,同时获得了仪表检测光强和细菌浓度关系曲线。针对实际样品检测中存在的问题,提出了相应的数据处理改进算法,引入了校准方程修正系数,用于食品及卡介苗样品检测,获得了良好的检测结果。对猴头菇类食品样品检测的结果与培养计数法检测结果的R达到0.993,重复性检测的相对平均偏差(R.A.D.)为7.69 %,变异系数(CV)为9.07 %。对卡介苗(BCG)样品检测结果与培养计数法检测结果的R达到0.997,R.A.D.为5.72 %,CV为7.73 %。与传统培养计数法相比,该仪表具有较好的检测速度,准确度和重复性。
Abstract
A rapid detecting instrument of bacteria was presented based on the principle of Adenosine Triphosphate(ATP) bioluminescence.In the instrument, a high-precision A/D converter and a small Photomultiplier Tube (PMT) module were included to improve the optical detection sensitivity, and the optical, circuit and software designs were integrated to construct the rapid detecting system for bacteria combinated with homemade ATP bioluminescence biosensors.Standard bacterium suspensions were detected to study the relation of Relative Light Units (RLU) and the concentration of bacteria, and the Correlation Coefficient (R) is 0.976 0.In order to detect actual samples, an improved data processing algorithm was bring forward, and modified modulus were imported in the equation.Furthermore,the detecting concentration of bacteria in a hericium erinaceus sample was demonstrated by the proposed system, results show that the R is 0.993 compared with the result obtained by the standard Aerobic Plate Count (APC) method.The reproducibility of the system was tested and its Relative Average Deviation (R.A.D.) is 7.69 % and Coefficient of Variability (CV) is 9.07 %.The BCG vaccines were also detected by this system and the R is 0.997 compared with the result obtained by the APC method.its R.A.D.is 7.69 % and CV is 9.07 %.It is concluded that this system can complete the process of detection in few minutes, and can offer its high sensitivity and reproducibility.
田青, 罗金平, 刘晓红, 高从军, 刘春秀, 蔡新霞. 生物发光法细菌快速检测仪的研制及应用[J]. 光学 精密工程, 2010, 18(4): 771. TIAN Qing, LUO Jin-ping, LIU Xiao-hong, GAO Cong-jun, LIU Chun-xiu, CAI Xin-xia. Development and application of rapid detecting instrument for bacteria based on bioluminescence[J]. Optics and Precision Engineering, 2010, 18(4): 771.