Assessment of sample collection and storage methods for multicenter immunologic research in children.

Author(s): Matheson LA, Duong TT, Rosenberg AM, Yeung RS

Publication: J Immunol Methods, 2008, Vol. 339, Page 82-9

PubMed ID: 18771669 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 collection tube, and storage and shipping temperature on the recovery of recombinant TNF-alpha and IL-2 from spiked plasma. Aliquots of lithium heparin plasma were frozen at -80 degrees C for 4 days and shipped overnight on dry ice or stored for 4 days at 4 degrees C and then shipped overnight at ambient temperatures along with the specimens in the P100 tubes from which the plasma was separated upon arrival. After shipping, all specimens were stored at -80 degrees C.

   Summary of Findings:

   TNF-alpha recovery from spiked lithium-heparin plasma was low (<15%), regardless of storage/shipping conditions, but when blood collected in a P100 tube was spiked with TNF-alpha, stored for 4 days at 4 degrees C, and then shipped at ambient temperatures before separation at the receiving lab, more than 80% of the TNF-alpha was recovered. In contrast, IL-2 recovery was not significantly different between lithium-heparin plasma and specimens collected and shipped in P100 tubes, regardless of storage and shipping conditions (60-80%).

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Plasma

   Preservative Types

   • Frozen

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Protein	ELISA

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage temperature	4 degrees C -80 degrees C
   Storage	Between site transportation method	Mailed
   Storage	Specimen transport duration/condition	On dry ice At ambient temperatures
   Biospecimen Acquisition	Type of collection container/solution	P100 Lithium-heparin vacutainers

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2. Study Purpose

   The purpose of this study was to determine the effects of storage and shipping temperature and extraction method on RNA yield, integrity and amplification success. Blood specimens in Tempus and PAXgene tubes were frozen at -80 degrees C for 5 days and shipped overnight on dry ice or stored for 5 days at 4 degrees C and then shipped overnight at ambient temperatures. Specimens shipped on dry ice were stored at -80 degrees C upon arrival, but specimens shipped at ambient temperatures were stored at room temperature after arrival.

   Summary of Findings:

   The RNA yield was highest when blood was collected in Tempus tubes, stored at 4 degrees C, shipped at ambient temperatures and isolated using the Tempus kit (13.79 ug) and lowest (3.43 ug) when blood was collected and isolated using the Paxgene system. The yield and integrity of RNA isolated using Paxgene were similar in specimens stored at 4 degrees C and shipped at ambient temperatures and specimens stored at -80 degrees C and shipped on dry ice. Some specimens collected and isolated using Tempus showed a slight decrease in RNA integrity, but Beta-actin amplification was possible from all specimens, and band intensities reflected the RNA yield. The highest CD4/GAPDH ratio was detected in specimens collected in Tempus tubes, stored at 4 degrees C and shipped at ambient temperatures (3.04) and the lowest in specimens collected in Paxgene tubes, stored at 4 degrees C and shipped at ambient temperatures (1.70), or stored at -80 and shipped on dry ice (1.86).

   Biospecimens

   • Bodily Fluid - Blood

   Preservative Types

   • Other Preservative
   • PAXgene

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   RNA	Spectrophotometry
   RNA	Electrophoresis
   RNA	Real-time qRT-PCR
   RNA	RT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage temperature	-80 degrees C 4 degrees C
   Storage	Between site transportation method	Mailed
   Storage	Specimen transport duration/condition	On dry ice At ambient temperature
   Biospecimen Preservation	Type of fixation/preservation	Tempus PAXgene
   RT-PCR Specific	Targeted nucleic acid	Beta-actin
   Real-time qRT-PCR Specific	Targeted nucleic acid	CD-4 GAPDH
   Analyte Extraction and Purification	Analyte isolation method	Tempus spin RNA kit Paxgene blood RNA kit

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