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Gold nanoparticles (AuNP)-based aptasensor for enteropathogenic Escherichia coli detection

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Abstract

Background

Diarrhea is a major cause of severe gastrointestinal illness in the infant especially in many developing countries. Although this molecular technique has been accepted as standard technique to detect Diarrhea-causing EPEC, the practical aspect of this technique for in-site rapid screening purposes is still facing a major challenge. In this study, we characterized EPEC specific aptamers and applied it as an AuNP-based aptasensor for point of care (POC) diagnosis purpose.

Methods

As many as six selected DNA aptamers was screened using target bacteria and the bound aptamer was measured by qPCR technique. Moreover, Kd value for each optimal bound aptamer was measured by using the same technique. Colorimetry assay was applied to test specificity and LOD of AuNP-based aptasensor.

Results

Two DNA aptamers have been successfully obtained to detect Enteropathogenic Escherichia coli K.1.1. DNA aptamer S8-7 exhibited constant dissociation (Kd) value of 17.08 nM, while DNA aptamer S10-5 exhibited Kd value of 34.14 nM. AuNP-based aptasensor showed high selectivity and specificity for EPEC K.1.1 with a limit of detection (LOD) value of 105 CFU/mL. Truncation study on DNA aptamer S8-7 showed that elimination of primer binding sequence only slightly increased both performance of detection and LOD value of AuNP-based aptasensor.

Conclusion

Further study is necessary to improve AuNP-aptasensor performance such as through mutagenesis approach on targeted DNA aptamers before AuNP-based aptasensor can be applied as a biosensor in point of care (POC) diagnosis.

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Acknowledgements

This research was fully funded by Prioritas Nasional (PN) Obat 2018-2019. All facilities were supported by Research Center for Genetic Engineering, National Research and Innovation Agency (BRIN), Indonesia. We would like to thank to Prof. Dr.dr. Sri Budiarti from IPB University, Bogor who kindly provides the EPEC K.1.1. The authors would like to thank to Lita Meilina from Research Center for Genetic Engineering, National Research and Innovation Agency (BRIN) for her assistance in manuscript preparation.

Funding

This study was fully funded and supported by Research Center for Genetic Engineering, National Research and Innovation Agency, Indonesia.

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Authors and Affiliations

  1. Research Center for Genetic Engineering, National Research and Innovation Agency (BRIN), Raya Bogor Street KM.46, Cibinong, Bogor, West Java, 16911, Indonesia

    Bugi Ratno Budiarto, Apon Zaenal Mustopa & Ratih Asmana Ningrum

  2. Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Indonesia, Depok, 16424, Indonesia

    Nurul Amilia & Endang Saepudin

  3. Department of Chemistry, Faculty of Science and Technology, UIN Syarif Hidayatullah Jakarta, South Tangerang, 15412, Indonesia

    Nurul Amilia

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  1. Bugi Ratno Budiarto
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Correspondence to Apon Zaenal Mustopa.

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The authors declare the competing material interests of EPEC K.1.1 as a target strain for DNA aptamer was provided by Prof. Dr. dr. Sri Budiarti from IPB University.

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Budiarto, B.R., Mustopa, A.Z., Ningrum, R.A. et al. Gold nanoparticles (AuNP)-based aptasensor for enteropathogenic Escherichia coli detection. Mol Biol Rep 49, 9355–9363 (2022). https://doi.org/10.1007/s11033-022-07786-3

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  • DOI: https://doi.org/10.1007/s11033-022-07786-3

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