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Direct Detection of Extracellular Vesicle miRNAs Using a Single-Step RT-qPCR Assay

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Extracellular Vesicles in Diagnosis and Therapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2504))

Abstract

Extracellular vesicles (EVs) are biological carriers, and EV-associated miRNAs (EV-miRNAs) are considered as a novel biomarker in multiple diseases. Currently, the column-based purification method is used to purify miRNAs from EVs. However, this method of purification is complex, time-consuming, and expensive. Therefore, a simple and cost-effective single-step quantitative reverse transcription-polymerase chain reaction (RT-qPCR) method is required to detect the expression of EV-miRNAs. This chapter describes a protocol for directly analyzing the EV-miRNAs expression from mouse bronchoalveolar lavage fluid (BALF) and serum without going for an RNA isolation and purification step from EVs. It is an efficient method in several terms such as cost-wise, time, low expertise, and accuracy in results. This method may be helpful in diagnostic blood tests used in medical centers or research laboratories.

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Author information

Authors and Affiliations

  1. Division of Pulmonary and Critical Care Medicine, Department of Medicine, Boston University, Boston, MA, USA

    Ayyanar Sivanantham & Yang Jin

  2. Department of Biology and Chemistry, Changwon National University, Changwon, Korea

    Heedoo Lee

Authors
  1. Ayyanar Sivanantham
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  2. Heedoo Lee
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  3. Yang Jin
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Corresponding author

Correspondence to Yang Jin .

Editor information

Editors and Affiliations

  1. National Center for Global Health, Istituto Superiore di SanitĂ , Rome, Italy

    Maurizio Federico

  2. National Center for Global Health, Istituto Superiore di SanitĂ , Rome, Italy

    Barbara Ridolfi

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Sivanantham, A., Lee, H., Jin, Y. (2022). Direct Detection of Extracellular Vesicle miRNAs Using a Single-Step RT-qPCR Assay. In: Federico, M., Ridolfi, B. (eds) Extracellular Vesicles in Diagnosis and Therapy. Methods in Molecular Biology, vol 2504. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2341-1_10

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  • DOI: https://doi.org/10.1007/978-1-0716-2341-1_10

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2340-4

  • Online ISBN: 978-1-0716-2341-1

  • eBook Packages: Springer Protocols

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