Examination of stability of bone marrow blood RNA in the PAXgene tube.

Author(s): Yamamoto T, Sekiyama A, Sekiguchi H, Yoshida T, Miyagi Y

Publication: Lab Hematol, 2006, Vol. 12, Page 143-7

PubMed ID: 16950675 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 preservation with PAXgene and refrigerated storage on RNA yield, integrity, and amplification success of bone marrow blood specimens from leukemia patients in remission. Bone marrow blood was collected from 9 individuals into EDTA tubes, and blood cells were transferred immediately to EDTA and PAXgene tubes and stored at 4 degrees C until analysis on days 0, 3, and 5. RNA extraction was with trizol.

   Summary of Findings:

   RNA yield from bone marrow blood decreased with increasing refrigerated storage in EDTA or PAXgene tubes. On day 0, the RNA yield was lower from PAXgene specimens than EDTA specimens and the decrease in RNA yield was much faster between days 0 and 3 when stored in PAXgene tubes rather than EDTA tubes. After day 3, the rate of decrease in RNA yield in PAXgene tubes was slightly less than that in EDTA tubes. In specimens from a single patient, the 28S rRNA band was of lower intensity in the PAXgene specimens stored 3 or 5 days than any of the EDTA specimens, regardless of storage. Further, the intensity tended to decrease for PAXgene and EDTA specimens with increasing storage time. In contrast, the intensity of the 18S rRNA band was higher in the PAXgene specimens stored for 3 or 5 days than in any EDTA specimens, regardless of storage time. On average, bone marrow blood stored in PAXgene tubes for 3 or 5 days had a higher concentration of PBGD transcripts than unstored bone marrow blood in EDTA tubes. Further, BCR-ABL was detected at low levels in specimens from a single patient stored in PAXgene but was not detected in specimens stored in EDTA tubes. In contrast, average Wilm's tumor suppressor WT1 levels were lower in PAXgene specimens stored 3 or 5 days than in unstored EDTA specimens, but levels remained relatively stable with storage.

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Bone Marrow
   • Cell - Blood

   Preservative Types

   • None (Fresh)
   • PAXgene

   Diagnoses:

   • Neoplastic - Leukemia

   Platform:

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

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Refrigeration None (fresh) PAXgene
   Storage	Storage duration	0 days 3 days 5 days
   Real-time qRT-PCR Specific	Targeted nucleic acid	BCR-ABL fusion PBGD WT1

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