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

Animals Biological Assay Biomarkers / metabolism Extracellular Vesicles / genetics Mice MicroRNAs / metabolism Real-Time Polymerase Chain Reaction
Bronchoalveolar lavage fluid (BALF) Cost-effective method EV-miRNAs Extracellular vesicles (EVs) Serum Single-step RT-qPCR

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2022
Historique:
entrez: 25 4 2022
pubmed: 26 4 2022
medline: 28 4 2022
Statut: ppublish

Résumé

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.

Identifiants

DOI: 10.1007/978-1-0716-2341-1_10 PMID: 35467284
pubmed: 35467284
doi: 10.1007/978-1-0716-2341-1_10
doi:

Substances chimiques

Biomarkers 0
MicroRNAs 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

137-145

Subventions

Organisme : NHLBI NIH HHS
ID : R01 HL142758
Pays : United States
Organisme : NIAID NIH HHS
ID : R33 AI121644
Pays : United States

Informations de copyright

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

Références

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Auteurs

Ayyanar Sivanantham (A)

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

Heedoo Lee (H)

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

Yang Jin (Y)

Division of Pulmonary and Critical Care Medicine, Department of Medicine, Boston University, Boston, MA, USA. yjin1@bu.edu.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › Direct Detection of Extracellular Vesicle...
questionsmedicales.fr › Techniques d'investigation › Dosage biologique › Direct Detection of Extracellular Vesicle...
questionsmedicales.fr › Facteurs biologiques › Marqueurs biologiques › Direct Detection of Extracellular Vesicle...
questionsmedicales.fr › Cellules › Structures cellulaires › Espace extracellulaire › Vésicules extracellulaires › Direct Detection of Extracellular Vesicle...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Rodentia › Muridae › Murinae › Souris › Direct Detection of Extracellular Vesicle...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN non traduit › Petit ARN non traduit › microARN › Direct Detection of Extracellular Vesicle...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Techniques d'amplification d'acides nucléiques › Réaction de polymérisation en chaîne › Réaction de polymérisation en chaine en temps réel › Direct Detection of Extracellular Vesicle...