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, and Doolan, Denise L. (2022) The Effect of Tropical Temperatures on the Quality of RNA Extracted from Stabilized Whole-Blood Samples. International Journal of Molecular Sciences, 23 (18). 10609.
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Abstract
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 RNAsequencing (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.
| Item ID: | 77199 |
|---|---|
| Item Type: | Article (Research - C1) |
| ISSN: | 1422-0067 |
| Related URLs: | |
| Copyright Information: | © 2022 by the authors. 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/). |
| Funders: | National Health and Medical Research Council of Australia (NHMRC) |
| Projects and Grants: | NHMRC #1069466, NHMRC #1137285 |
| Date Deposited: | 16 Jan 2023 22:15 |
| FoR Codes: | 32 BIOMEDICAL AND CLINICAL SCIENCES > 3299 Other biomedical and clinical sciences > 329999 Other biomedical and clinical sciences not elsewhere classified @ 100% |
| SEO Codes: | 20 HEALTH > 2001 Clinical health > 200199 Clinical health not elsewhere classified @ 100% |
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