Cargando…

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...

Descripción completa

Detalles Bibliográficos
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
Descripción
Sumario: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.