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Colorimetric assessment of BCR-ABL1 transcripts in clinical samples via gold nanoprobes

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Abstract

Gold nanoparticles functionalized with thiolated oligonucleotides (Au-nanoprobes) have been used in a range of applications for the detection of bioanalytes of interest, from ions to proteins and DNA targets. These detection strategies are based on the unique optical properties of gold nanoparticles, in particular, the intense color that is subject to modulation by modification of the medium dieletric. Au-nanoprobes have been applied for the detection and characterization of specific DNA sequences of interest, namely pathogens and disease biomarkers. Nevertheless, despite its relevance, only a few reports exist on the detection of RNA targets. Among these strategies, the colorimetric detection of DNA has been proven to work for several different targets in controlled samples but demonstration in real clinical bioanalysis has been elusive. Here, we used a colorimetric method based on Au-nanoprobes for the direct detection of the e14a2 BCR-ABL fusion transcript in myeloid leukemia patient samples without the need for retro-transcription. Au-nanoprobes directly assessed total RNA from 38 clinical samples, and results were validated against reverse transcription-nested polymerase chain reaction (RT-nested PCR) and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The colorimetric Au-nanoprobe assay is a simple yet reliable strategy to scrutinize myeloid leukemia patients at diagnosis and evaluate progression, with obvious advantages in terms of time and cost, particularly in low- to medium-income countries where molecular screening is not routinely feasible.

Gold nanoprobe for colorimetric detection of BCR-ABL1 fusion transcripts originating from the Philadelphia chromosome

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Acknowledgments

We acknowledge Fundação para a Ciência e a Tecnologia (FCT/MEC) for financial support (Project PTDC/BBB-NAN/1812/2012 and UCIBIO UID/Multi/04378/2013) and co-financed by ERDF under PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728), RV for SFRH/BD/52211/2013.

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Authors and Affiliations

  1. UCIBIO, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus da Caparica, 2829-516, Caparica, Portugal

    Raquel Vinhas, Cláudia Correia, Alexandra R. Fernandes & Pedro V. Baptista

  2. Serviço de Hematologia, Hospital dos Capuchos (CHLC), 1169-050, Lisbon, Portugal

    Patricia Ribeiro, Alexandra Lourenço & Aida Botelho de Sousa

Authors
  1. Raquel Vinhas
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  2. Cláudia Correia
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  3. Patricia Ribeiro
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  4. Alexandra Lourenço
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  5. Aida Botelho de Sousa
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  6. Alexandra R. Fernandes
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  7. Pedro V. Baptista
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Corresponding author

Correspondence to Pedro V. Baptista.

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The authors declare that they have no competing interests.

Informed consent

Patients were sent to the hematology department of the Hospital dos Capuchos (CHLC, Lisbon, Portugal) for leukemia diagnosis. Written informed consent was obtained from all participants and the study was approved by the Hospital dos Capuchos Ethics Committee. All approved ethical requirements for sample collection and assortment, processing, and analysis required by the Hospital dos Capuchos Ethics Committee have been strictly followed.

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Vinhas, R., Correia, C., Ribeiro, P. et al. Colorimetric assessment of BCR-ABL1 transcripts in clinical samples via gold nanoprobes. Anal Bioanal Chem 408, 5277–5284 (2016). https://doi.org/10.1007/s00216-016-9622-9

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  • DOI: https://doi.org/10.1007/s00216-016-9622-9

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