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Conductive silver paste smeared glass substrates for label-free Raman spectroscopic detection of HIV-1 and HIV-1 p24 antigen in blood plasma

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Abstract

We report on application of conductive silver paste smeared glass slides as Raman spectroscopy sample substrates for label-free detection of HIV-1 p24 antigen in blood plasma. We also show that the same substrates can be applied in Raman spectroscopic screening of blood plasma for presence of HIV. The characteristic Raman spectrum of HIV-1 p24 antigen displayed prominent bands that were assigned to ribonucleic acids (RNA) and proteins that constitute the antigen. This spectrum can be used as reference during Raman spectroscopic screening for HIV in plasma within the first few days after exposure (<7 days). The Raman spectra obtained from HIV+ plasma displayed unique peaks centered at wavenumbers 928, 990, 1270, 1397, and 1446 cm−1 attributed to the Raman active vibrations in the virion carbohydrates, lipids, and proteins. Other bands similar to those reported in literature were also seen and assignments made. The attachment of the HIV virions to silver nanoparticles via gp120 glycoprotein knobs was thought to be responsible for the enhanced Raman signals of proteins associated with the virus. The principal component analysis (PCA) applied on the combined spectral data showed that HIV− and HIV+ spectra had differing spectral patterns. This indicated the great power of Raman spectroscopy in HIV detection when plasma samples are deposited onto silver paste smeared glass substrates. The Raman peaks responsible for the segregation of the spectral data in PCA were mainly those assigned to the viral proteins (645, 725, 813, 1270, and 1658 cm−1). Excellent results were obtained from Artificial Neural Network (ANN) applied on the HIV+ Raman spectral data around the prominent peak centered at 1270 cm−1 with R (coefficient of correlation) and R 2 (coefficient of determination) values of 0.9958 and 0.9895, respectively. The method has the potential of being used as quick blood screening for HIV before blood transfusion with the Raman peaks assigned to the virion proteins acting as reference.

The HIV type 1 virus particle gets attached to the silver nanoparticle contained in the conductive silver paste smear onto a glass slide. This results in strong Raman signals associated with the components of the virion. The signals are collected, dispersed in a spectrometer and displayed on a computer screen. Method can be used as a label-free and rapid HIV screening in blood plasma

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Acknowledgements

The authors would like to thank Dr. Julius O. Oyugi of Department of Medical Microbiology, University of Nairobi, for providing the samples used in the study. The first author thanks NACOSTI, Kenya, for funding his studies. We also thank ISP Uppsala University for grant KEN004.

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Authors and Affiliations

  1. Department of Physics, Egerton University, P.O Box 536-20115, Egerton, 20115, Kenya

    Ben O. Otange & Ronald Rop

  2. Department of Physics, University of Nairobi, P.O Box 30197-00100, Nairobi, 00100, Kenya

    Zephania Birech & Justus Okonda

Authors
  1. Ben O. Otange
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  2. Zephania Birech
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  3. Justus Okonda
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  4. Ronald Rop
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Correspondence to Zephania Birech.

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This study was approved by the Ethics and Research Committee of the University of Nairobi (KNH-UoN:P637=10=2015).

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The authors declare that they have no competing interest.

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Otange, B.O., Birech, Z., Okonda, J. et al. Conductive silver paste smeared glass substrates for label-free Raman spectroscopic detection of HIV-1 and HIV-1 p24 antigen in blood plasma. Anal Bioanal Chem 409, 3253–3259 (2017). https://doi.org/10.1007/s00216-017-0267-0

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  • DOI: https://doi.org/10.1007/s00216-017-0267-0

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