Journal of Applied Optics, Volume. 44, Issue 4, 859(2023)

Multiple real-time fluorescence detection system for rapid on-site nucleic acid detection

Jiayu YANG1...2,3,4, Juntian ZENG1,2,3,4, Bangchao XI1,2,3,4, Guoqiang LIU1,2,3,4, Shaolei HUANG1,2,3,4, and Dongxu ZHANG1,2,34,* |Show fewer author(s)
Author Affiliations
  • 1School of Public Health, Xiamen University, Xiamen 361102, China
  • 2State Key Laboratory of Molecular Vaccinology and Molecular Diagnostic, Xiamen University, Xiamen 361102, China
  • 3National Institute of Diagnostics and Vaccine Development in Infection Diseases, Xiamen University, Xiamen 361102, China
  • 4State Drug Administration Key Laboratory of Infectious Disease Detection Technology Research and Evaluation, Xiamen 361102, China
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    Figures & Tables(17)
    Excitation optical path and emission optical path
    Semi-divergence angle test device
    Comparison diagram of emission optical path simulation and illumination
    Multi-channel rotation integrated module
    Embedded structure between module brackets
    Physical image of proposed fluorescence detection system
    Test results of fluorescence dye gradient for each channel
    Test results of fluorescence crosstalk between channels
    Test results of sample amplification detection
    • Table 1. Semi-divergence angle measured at each end of optical path(°)

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      Table 1. Semi-divergence angle measured at each end of optical path(°)

      位置激发光路前端芯片激发端发射光路前端
      发散角19.0313.5018.78
    • Table 2. Distance between elements of emission optical pathmm

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      Table 2. Distance between elements of emission optical pathmm

      距离发射光纤-透镜透镜-发射滤光片发射滤光片-PD
      优化前距离11.83.73.1
      优化后距离8.85.82.5
    • Table 3. Dilution concentration of fluorescent dyesμg·mL−1

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      Table 3. Dilution concentration of fluorescent dyesμg·mL−1

      荧光染料FAMHEXROXCY5
      母液(50%DMSO)2 0001320018
      梯度1(DEPC水)2001.3201.8
      梯度2(DEPC水)1000.65100.9
      梯度3(DEPC水)500.32550.45
      梯度4(DEPC水)250.162 52.50.225
      梯度5(DEPC水)12.50.081 251.250.112 5
    • Table 4. Detection limit test results of fluorescent dyes

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      Table 4. Detection limit test results of fluorescent dyes

      检测通道ROXFAMHEXCY5
      检测下限/μg·mL−10.311.560.020.11
      线性度R20.99610.94720.99680.9935
    • Table 5. Gradient test results of fluorescent dyes

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      Table 5. Gradient test results of fluorescent dyes

      检测通道电流值对数线性度R2
      梯度5梯度4梯度3梯度2梯度1
      ROX2.993.884.845.886.840.999 3
      FAM1.702.213.224.175.010.990 7
      HEX5.676.547.438.619.390.996 5
      CY54.615.276.106.877.390.994 8
    • Table 6. Repeatability test results of fluorescence intensity

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      Table 6. Repeatability test results of fluorescence intensity

      检测通道CV值
      梯度5/%梯度3/%梯度1/%
      ROX1.270.190.16
      FAM1.200.890.59
      HEX1.010.270.06
      CY51.240.200.29
    • Table 7. Fluorescence crosstalk test results based on HEX fluorescent dyespA

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      Table 7. Fluorescence crosstalk test results based on HEX fluorescent dyespA

      检测通道电流值
      高浓度中浓度低浓度DEPC水
      ROX1.901.112.151.28
      FAM0.290.541.230.22
      HEX217.7165.6320.524.18
      CY50.800.420.671.19
    • Table 8. Fluorescence crosstalk test results based on CY5 fluorescent dyespA

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      Table 8. Fluorescence crosstalk test results based on CY5 fluorescent dyespA

      检测通道电流值
      高浓度中浓度低浓度DEPC水
      ROX0.220.370.671.15
      FAM0.280.140.190.02
      HEX2.030.191.781.37
      CY5120.6338.5414.150.94
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    Jiayu YANG, Juntian ZENG, Bangchao XI, Guoqiang LIU, Shaolei HUANG, Dongxu ZHANG. Multiple real-time fluorescence detection system for rapid on-site nucleic acid detection[J]. Journal of Applied Optics, 2023, 44(4): 859

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    Paper Information

    Category: Research Articles

    Received: Sep. 29, 2022

    Accepted: --

    Published Online: Aug. 10, 2023

    The Author Email:

    DOI:10.5768/JAO202344.0403007

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