Human blood RNA stabilization in samples collected and transported for a large biobank.

Author(s): Duale N, Brunborg G, Rønningen KS, Briese T, Aarem J, Aas KK, Magnus P, Stoltenberg C, Susser E, Lipkin WI

Publication: BMC Res Notes, 2012, Vol. 5, Page 510

PubMed ID: 22988904 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of storage duration and RNA stabilization system on levels of RNA in adult blood and cord blood. Blood was frozen in collection tubes at -20 degrees C and then stored at -80 degrees C until RNA extraction.

   Summary of Findings:

   RNA yields were 2-3 fold higher when blood was collected and stored in Tempus tubes rather than PAXgene tubes (p<0.05). While RIN and OD at 260/280 ratio were comparable for PAXgene and Tempus specimens, the OD 260/230 ratio was lower for RNA from PAXgene tubes (p<0.05). RNA quality was not affected by storage of the blood at room temperature for up to 7 days, but RNA yield was slightly lower when specimens were stored in PAXgene tubes for 7 days then when RNA was extracted on day 0, and RNA transcript levels were affected by storage at room temperature for 2 days or more. The effects of room temperature storage on levels of RNA transcripts were more pronounced in specimens stored in PAXgene tubes than Tempus tubes, but all changes were less than 2 fold. RNA yield was higher from umbilical cord blood than adult blood.

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Cord Blood

   Preservative Types

   • Other Preservative
   • Frozen
   • PAXgene

   Diagnoses:

   • Not specified
   • Pregnant

   Platform:

   Analyte	Technology Platform
   RNA	Automated electrophoresis/Bioanalyzer
   RNA	Real-time qRT-PCR
   RNA	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Frozen PAXgene Tempus
   Biospecimen Aliquots and Components	Blood and blood products	Umbilical cord blood Whole blood
   Biospecimen Acquisition	Type of collection container/solution	Tempus tube PAXgene tube
   Storage	Time at room temperature	0 days 2 days 5 days 7 days
   Real-time qRT-PCR Specific	Targeted nucleic acid	Cyclin-dependent kinase inhibitor (CDKN1A) Interleukin 8 (IL8) MYC Tumor protein 53 (TP53)

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2. Study Purpose

   The purpose of this study was to determine the effects of transport distance and temperature on levels of RNA in adult blood and cord blood. All specimens were stored in Tempus collection tubes at room temperature until arrival, at which point, all blood was frozen in collection tubes at -20 degrees C at the same time and then stored at -80 degrees C until RNA extraction. Transport temperatures ranged from 3.4-27.1 degrees C.

   Summary of Findings:

   RNA quality, as determined by RIN, decreased when specimens were transported (p<0.05), but the magnitude of the decrease depended on transport duration and temperature which were each dependent on the destination. The specimens least affected by transport were mailed from Oslo-Baerum and experienced lower temperatures (<6 degrees C) for two 12 h periods during transport. Relative transcript levels were also affected by transport duration and temperature. There was no effect of transport on OD 260/280 ratios or OD 260/230 ratios.

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Cord Blood

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Not specified
   • Pregnant

   Platform:

   Analyte	Technology Platform
   RNA	Automated electrophoresis/Bioanalyzer
   RNA	Real-time qRT-PCR
   RNA	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Between site transportation method	Mailed Not transported
   Storage	Specimen transport duration/condition	Mailed between Oslo and Kirkenes Norway Mailed between Oslo and Bergen Norway Mailed between Oslo and Kristians Norway Mailed between Oslo and Baerum Norway
   Real-time qRT-PCR Specific	Targeted nucleic acid	CDKN1A FOS IL8 MYC IL1B TP53

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3. Study Purpose

   The purpose of this study was to determine the effects of blood collection volume on levels of RNA in umbilical cord and adult blood. The recommended collection volume for the Tempus tubes was 3.0 mL.

   Summary of Findings:

   RNA yield, RIN, OD 260/280 and OD 260/230 ratios decreased with decreasing blood volume (p<0.05, all), but effects on RIN, OD 260/280 and OD 260/230 were only observed when blood volumes of less than 2 mL adult blood or 0.5 mL umbilical cord blood were compared to a 3.0 mL collection volume. Compared to the 3.0 mL collection volume, relative transcript levels were affected by the collection of less than 1.5 mL adult blood, less than 1.0 mL umbilical cord blood or by collection of more than 3.0 mL of either type of blood.

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Cord Blood

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Not specified
   • Pregnant

   Platform:

   Analyte	Technology Platform
   RNA	Automated electrophoresis/Bioanalyzer
   RNA	Real-time qRT-PCR
   RNA	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Blood and blood products	Umbilical cord blood Whole blood
   Biospecimen Aliquots and Components	Aliquot size/volume	0.5 mL 1.0 mL 1.5 mL 2.0 mL 3.0 mL 3.3 mL
   Real-time qRT-PCR Specific	Targeted nucleic acid	CDKN1A FOS IL8 MYC IL1B TP53

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