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The Effect of Tropical Temperatures on the Quality of RNA Extracted from Stabilized Whole-Blood Samples

Whole-blood-derived transcriptional profiling is widely used in biomarker discovery, immunological research, and therapeutic development. Traditional molecular and high-throughput transcriptomic platforms, including molecular assays with quantitative PCR (qPCR) and RNA-sequencing (RNA-seq), are depe...

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Autores principales: Sarathkumara, Yomani D., Browne, Daniel J., Kelly, Ashton M., Pattinson, David J., Rush, Catherine M., Warner, Jeffrey, Proietti, Carla, Doolan, Denise L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9503649/
https://www.ncbi.nlm.nih.gov/pubmed/36142559
http://dx.doi.org/10.3390/ijms231810609
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author Sarathkumara, Yomani D.
Browne, Daniel J.
Kelly, Ashton M.
Pattinson, David J.
Rush, Catherine M.
Warner, Jeffrey
Proietti, Carla
Doolan, Denise L.
author_facet Sarathkumara, Yomani D.
Browne, Daniel J.
Kelly, Ashton M.
Pattinson, David J.
Rush, Catherine M.
Warner, Jeffrey
Proietti, Carla
Doolan, Denise L.
author_sort Sarathkumara, Yomani D.
collection PubMed
description Whole-blood-derived transcriptional profiling is widely used in biomarker discovery, immunological research, and therapeutic development. Traditional molecular and high-throughput transcriptomic platforms, including molecular assays with quantitative PCR (qPCR) and RNA-sequencing (RNA-seq), are dependent upon high-quality and intact RNA. However, collecting high-quality RNA from field studies in remote tropical locations can be challenging due to resource restrictions and logistics of post-collection processing. The current study tested the relative performance of the two most widely used whole-blood RNA collection systems, PAXgene(®) and Tempus™, in optimal laboratory conditions as well as suboptimal conditions in tropical field sites, including the effects of extended storage times and high storage temperatures. We found that Tempus™ tubes maintained a slightly higher RNA quantity and integrity relative to PAXgene(®) tubes at suboptimal tropical conditions. Both PAXgene(®) and Tempus™ tubes gave similar RNA purity (A260/A280). Additionally, Tempus(™) tubes preferentially maintained the stability of mRNA transcripts for two reference genes tested, Succinate dehydrogenase complex, subunit A (SDHA) and TATA-box-binding protein (TBP), even when RNA quality decreased with storage length and temperature. Both tube types preserved the rRNA transcript 18S ribosomal RNA (18S) equally. Our results suggest that Tempus(™) blood RNA collection tubes are preferable to PAXgene(®) for whole-blood collection in suboptimal tropical conditions for RNA-based studies in resource-limited settings.
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spelling pubmed-95036492022-09-24 The Effect of Tropical Temperatures on the Quality of RNA Extracted from Stabilized Whole-Blood Samples Sarathkumara, Yomani D. Browne, Daniel J. Kelly, Ashton M. Pattinson, David J. Rush, Catherine M. Warner, Jeffrey Proietti, Carla Doolan, Denise L. Int J Mol Sci Article Whole-blood-derived transcriptional profiling is widely used in biomarker discovery, immunological research, and therapeutic development. Traditional molecular and high-throughput transcriptomic platforms, including molecular assays with quantitative PCR (qPCR) and RNA-sequencing (RNA-seq), are dependent upon high-quality and intact RNA. However, collecting high-quality RNA from field studies in remote tropical locations can be challenging due to resource restrictions and logistics of post-collection processing. The current study tested the relative performance of the two most widely used whole-blood RNA collection systems, PAXgene(®) and Tempus™, in optimal laboratory conditions as well as suboptimal conditions in tropical field sites, including the effects of extended storage times and high storage temperatures. We found that Tempus™ tubes maintained a slightly higher RNA quantity and integrity relative to PAXgene(®) tubes at suboptimal tropical conditions. Both PAXgene(®) and Tempus™ tubes gave similar RNA purity (A260/A280). Additionally, Tempus(™) tubes preferentially maintained the stability of mRNA transcripts for two reference genes tested, Succinate dehydrogenase complex, subunit A (SDHA) and TATA-box-binding protein (TBP), even when RNA quality decreased with storage length and temperature. Both tube types preserved the rRNA transcript 18S ribosomal RNA (18S) equally. Our results suggest that Tempus(™) blood RNA collection tubes are preferable to PAXgene(®) for whole-blood collection in suboptimal tropical conditions for RNA-based studies in resource-limited settings. MDPI 2022-09-13 /pmc/articles/PMC9503649/ /pubmed/36142559 http://dx.doi.org/10.3390/ijms231810609 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Sarathkumara, Yomani D.
Browne, Daniel J.
Kelly, Ashton M.
Pattinson, David J.
Rush, Catherine M.
Warner, Jeffrey
Proietti, Carla
Doolan, Denise L.
The Effect of Tropical Temperatures on the Quality of RNA Extracted from Stabilized Whole-Blood Samples
title The Effect of Tropical Temperatures on the Quality of RNA Extracted from Stabilized Whole-Blood Samples
title_full The Effect of Tropical Temperatures on the Quality of RNA Extracted from Stabilized Whole-Blood Samples
title_fullStr The Effect of Tropical Temperatures on the Quality of RNA Extracted from Stabilized Whole-Blood Samples
title_full_unstemmed The Effect of Tropical Temperatures on the Quality of RNA Extracted from Stabilized Whole-Blood Samples
title_short The Effect of Tropical Temperatures on the Quality of RNA Extracted from Stabilized Whole-Blood Samples
title_sort effect of tropical temperatures on the quality of rna extracted from stabilized whole-blood samples
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9503649/
https://www.ncbi.nlm.nih.gov/pubmed/36142559
http://dx.doi.org/10.3390/ijms231810609
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