Sensor Arrays for Electrochemical Detection of PCR-Amplified Genes Extracted from Cells Suspended in Environmental Waters

Ecological surveys of living things based on DNAs from environmental samples are attractive. However, despite simplicity of water sampling from the target environment, it is still necessary to transport the samples to the laboratory for DNA analysis based on skillful next-generation sequencers. To perform DNA-oriented surveys based on a simple protocol without any special training, we demonstrated, in this study, the detection of genes from cell-containing environmental waters using gene sensor arrays that require no DNA labeling and no external indicators. Cell-suspended PBS or river water were used as models of environmental waters containing living things, and DNA samples were prepared by PCR amplification. Ferrocene-terminated probes were synthesized and immobilized on an electrode array to develop a sensor array. The sensor array showed a large response to a target DNA complementary to the probe and no response to a mismatched DNA, indicating sequence-specific detection. For DNA samples prepared from the cells in PBS, they showed good responses similar to those for the target DNA. They also significantly detected DNA samples from the cells in river water at a general environmental concentration (38 cells mL<sup>-1</sup>) with 28-fold larger responses than those for 0 cells mL<sup>-1</sup>.

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Publication Details

Journal
Sensors
Published
2024-11-08
DOI
https://doi.org/10.3390/s24227182
Citations
2
Primary Topic
Environmental DNA in Biodiversity Studies
Type
article
Field-Weighted Citation Impact
0.92

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article

Sensor Arrays for Electrochemical Detection of PCR-Amplified Genes Extracted from Cells Suspended in Environmental Waters

Kazuki Miura, Mai Kawaguchi, Hiroshi Aoki, Siro Simizu et al.
2 citations
Sensors
Environmental DNA in Biodiversity Studies
0.92
article

Sensor Arrays for Electrochemical Detection of PCR-Amplified Genes Extracted from Cells Suspended in Environmental Waters

Kazuki Miura, Mai Kawaguchi, Hiroshi Aoki, Siro Simizu, Yuki Hiruta, Hiroki Kamo, Daniel Citterio, Yukiko Kumakura
article en
2 citations

Abstract

Ecological surveys of living things based on DNAs from environmental samples are attractive. However, despite simplicity of water sampling from the target environment, it is still necessary to transport the samples to the laboratory for DNA analysis based on skillful next-generation sequencers. To perform DNA-oriented surveys based on a simple protocol without any special training, we demonstrated, in this study, the detection of genes from cell-containing environmental waters using gene sensor arrays that require no DNA labeling and no external indicators. Cell-suspended PBS or river water were used as models of environmental waters containing living things, and DNA samples were prepared by PCR amplification. Ferrocene-terminated probes were synthesized and immobilized on an electrode array to develop a sensor array. The sensor array showed a large response to a target DNA complementary to the probe and no response to a mismatched DNA, indicating sequence-specific detection. For DNA samples prepared from the cells in PBS, they showed good responses similar to those for the target DNA. They also significantly detected DNA samples from the cells in river water at a general environmental concentration (38 cells mL<sup>-1</sup>) with 28-fold larger responses than those for 0 cells mL<sup>-1</sup>.

SensorsVol. 24(22)
Keio University (JP), National Institute of Advanced Industrial Science and Technology (JP)
Japan Society for the Promotion of Science
Openalex Percentile: Top 22%
Environmental DNA in Biodiversity Studies
0.92
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