Development and performance evaluation of flow cytometry for rapid quantitative detection of bacteria

Xin-tao YAN; Ce WANG; Zhi-guo PEI; Jue LIU

doi:10.3788/OPE.20192706.1245

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Development and performance evaluation of flow cytometry for rapid quantitative detection of bacteria

Life Science Instrument | 更新时间：2020-08-13

- Development and performance evaluation of flow cytometry for rapid quantitative detection of bacteria
- Optics and Precision Engineering Vol. 27, Issue 6, Pages: 1245-1254(2019)
- 作者机构：
  
  1.中国科学院苏州生物医学工程技术研究所中科院生物医学检验技术重点实验室，江苏苏州 215163
  2.复旦大学工程与应用技术研究院，上海 200433
  3.苏州高新区自来水有限公司，江苏苏州 215011
- 作者简介：
- 基金信息：
- DOI：10.3788/OPE.20192706.1245
  CLC： TH79; Q93-33
- Received：25 February 2019，
  
  Accepted：18 March 2019，
  
  Published：15 June 2019
- 稿件说明：
移动端阅览
Xin-tao YAN, Ce WANG, Zhi-guo PEI, et al. Development and performance evaluation of flow cytometry for rapid quantitative detection of bacteria[J]. Optics and precision engineering, 2019, 27(6): 1245-1254.
DOI：

Xin-tao YAN, Ce WANG, Zhi-guo PEI, et al. Development and performance evaluation of flow cytometry for rapid quantitative detection of bacteria[J]. Optics and precision engineering, 2019, 27(6): 1245-1254. DOI： 10.3788/OPE.20192706.1245.

摘要

为了实现对饮用水中细菌快速定量检测，建立了基于流式细胞术的高通量定量检测系统。对该系统的信号采集系统、绝对计数方法以及在细菌检测方面的综合性能进行了研究和评估。根据饮用水中典型细菌的荧光染料及其荧光激发光谱特点，介绍了流式细胞术快速检测细菌的工作原理及硬件平台。通过简化细菌的荧光信号强度计算模型，评估了信号采集系统的信噪比。建立了基于流量传感器的绝对计数方法，将检测系统与以参比微球法进行绝对计数的BD LSR Fortessa进行了一系列对比实验和统计学分析，测试和评估了检测系统在饮用水中细菌检测的综合性能水平。实验结果表明：对于4 MHz宽带的荧光信号，信号采集系统的信噪比可达86 dB；对于一定浓度内的微球，系统对它测试

值低于2%，与BD仪器测试结果的相关系数高达0.999 6，对等比例稀释的微球测试线性度高达0.999 8，最低可检测细菌浓度可达10

particles/mL；系统对E.coli和S.aureus含量测试结果的

值均低于7%

与BD仪器测试结果的相关系数均高于0.995 9，两仪器测试结果的相对误差均在4%以内。该仪器能实现对细菌的高精度快速定量检测，为饮用水中典型细菌快速检测仪器的开发提供了参考。

Abstract

To realize rapid quantitative detection of bacteria found in drinking water

a high-throughput quantitative detection system based on flow cytometry was established. The system's signal acquisition system

absolute counting method

and comprehensive performance for bacterial detection were investigated and evaluated. Firstly

according to the fluorescent dyes of typical bacteria found in drinking water and their fluorescence excitation spectra

the working principle and hardware platform of the flow cytometer for rapid bacterial detection were established. Secondly

the signal-to-noise ratio of the signal acquisition system was evaluated by simplifying the calculation model for signal strength of bacterial fluorescence. Next

a calculation method based on the flow sensor method was established for absolute counting. Finally

a series of statistically analyzed control experiments were performed with the self-developed instrument alongside BD LSR Fortessa with absolute reference according to the reference microsphere method. The performance of the self-developed instrument for the detection of bacteria in drinking water was comprehensively tested and evaluated. The experimental results indicated the followings: for a 4 MHz wideband fluorescent signal

the signal-to-noise ratio of the signal acquisition system can reach 86 dB; for microspheres in a certain concentration range

the

value of the system test results is less than 2%

the correlation coefficient with the test results of the BD instrument is as high as 0.999 6

the linearity of the equivalent dilution microsphere test is up to 0.999 8

and the lowest detectable bacterial concentration is 10

particles/mL; for

E.coli

and

S.aureus

bacteria

the

value of the test results of the system is less than 7%

the correlation coefficient with the test results of the BD instrument exceeds 0.995 9

and the relative deviation is within 4%. The instrument is capable of rapid quantitative detection of bacteria with a high-precision

providing a reference for the development of rapid detection instruments for typical bacteria found in drinking water.

关键词

Keywords

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