Variation of Peripheral Blood Mononuclear Cell RNA Quality in Archived Samples.

The Infectious Diseases BioBank (IDB) has consistently archived peripheral blood mononuclear cell (PBMNC) RNA for transcriptome analyses. RNA is particularly labile, and hence, these samples provide a sensitive indicator for assessing the IDB's quality-assurance measures. Independent analyses of 104 PBMNC RNA specimens from 26 volunteers revealed that the mean RNA integrity number (RIN) was high (9.02), although RIN ranged between scores of 7 and 10. This variation of RIN values was not associated with ischemic time, PBMNC quality, number of samples processed per day, self-medication after immunization, freezer location, donor characteristics, differential white blood cell counts, or daily variation in RNA extractions (all P>0.05). RIN values were related to the date of collection, with those processed during mid-summer having highest RIN scores (P=0.0001). Amongst specimens with the lowest RIN scores, no common feature could be identified. Thus, no technical explanation for the variation in RNA quality could be ascertained and these may represent normal physiological variations. These data provide strong evidence that current IDB protocols for the isolation and preservation PBMNC RNA are robust.

[1]  Fotini Betsou,et al.  Serum biobank certification and the establishment of quality controls for biological fluids: examples of serum biomarker stability after temperature variation , 2007, Clinical chemistry and laboratory medicine.

[2]  Karl Kornacker,et al.  Chipping away at the chip bias: RNA degradation in microarray analysis , 2003, Nature Genetics.

[3]  J. Bland The Physical and Chemical Properties of Ribosomes. , 1965 .

[4]  P. Elliott,et al.  Current standards for the storage of human samples in biobanks , 2010, Genome Medicine.

[5]  Thomas Ragg,et al.  The RIN: an RNA integrity number for assigning integrity values to RNA measurements , 2006, BMC Molecular Biology.

[6]  J. Cason,et al.  The Infectious Diseases BioBank at King's College London: archiving samples from patients infected with HIV to facilitate translational research , 2009, Retrovirology.

[7]  G. Ahmann,et al.  Effect of Tissue Shipping on Plasma Cell Isolation, Viability, and RNA Integrity in the Context of a Centralized Good Laboratory Practice–Certified Tissue Banking Facility , 2008, Cancer Epidemiology Biomarkers & Prevention.

[8]  D. Mizuno,et al.  Increase in the ratio of 18S RNA to 28S RNA in the cytoplasm of mouse tissues during aging , 1978, Mechanisms of Ageing and Development.

[9]  A. Reinberg,et al.  Seasonal modulation of the circadian time structure of circulating T and natural killer lymphocyte subsets from healthy subjects. , 1988, The Journal of clinical investigation.

[10]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[11]  Charles Auffray,et al.  Towards standardization of RNA quality assessment using user-independent classifiers of microcapillary electrophoresis traces , 2005, Nucleic acids research.

[12]  B. Jaber,et al.  Whole Blood Transcriptomics in Cardiac Surgery Identifies a Gene Regulatory Network Connecting Ischemia Reperfusion with Systemic Inflammation , 2010, PloS one.

[13]  Joakim Dillner,et al.  Ensuring quality in studies linking cancer registries and biobanks , 2010, Acta oncologica.

[14]  Stephen L. R. Ellison,et al.  Evaluation of a novel approach for the measurement of RNA quality , 2010, BMC Research Notes.

[15]  Mario Plebani,et al.  Post-analytical errors with portable glucose meters in the hospital setting. , 2009, Clinica chimica acta; international journal of clinical chemistry.

[16]  E. Thiel,et al.  Prediction of qualitative outcome of oligonucleotide microarray hybridization by measurement of RNA integrity using the 2100 Bioanalyzer™ capillary electrophoresis system , 2009, Annals of Hematology.

[17]  W. Gerald,et al.  Biobanking of Human Pancreas Cancer Tissue: Impact of Ex-Vivo Procurement Times on RNA Quality , 2010, Annals of Surgical Oncology.

[18]  Mario Plebani,et al.  Errors in a stat laboratory: types and frequencies 10 years later. , 2007, Clinical chemistry.

[19]  D. Whiteman,et al.  Serum Vitamin D Levels in Office Workers in a Subtropical Climate , 2011, Photochemistry and photobiology.