Blood Collection Tubes and Storage Temperature Should Be Evaluated when Using the Siemens ADVIA Centaur XP for Measuring 25-Hydroxyvitamin D

Abstract Background Efficient blood stabilization is essential to obtaining reliable and comparable RNA analysis data in preclinical operations. PAXgene (Qiagen, Becton Dickinson) and Tempus (Applied Biosystems, Life Technologies) blood collection tubes with RNA stabilizers both avoid preanalytical degradation of mRNA by endogenous nucleases and modifications in specific mRNA concentrations by unintentional up- or down-regulation of gene expression. Methods Sixteen different preanalytical conditions were tested in PAXgene and Tempus blood samples from seven donors: different mixing after collection, different fill volumes and different 24-h transport temperature conditions after collection. RNA was extracted by column-based methods. The quality of the extracted RNA was assessed by spectrophotometric quantification, A260/A280 purity ratio, RNA Integrity Number (Agilent Bioanalyzer), miRNA quantative real time polymerase chain reaction (qRT-PCR) on two target miRNAs (RNU-24 and miR-16), mRNA quality index by qRT-PCR on the 3′ and 5′ region of the GAPDH gene, and the PBMC preanalytical score, based on the relative expression levels of the IL8 and EDEM3 coding genes. Results When PAXgene RNA and Tempus blood collection tubes were used following the manufacturers’ instructions, there was no statistically or technically significant difference in the output RNA quality attributes. However, the integrity of the RNA extracted from Tempus collection tubes was more sensitive to fill volumes and effective inversion, than to storage temperature, while the integrity of RNA extracted from PAXgene collection tubes was more sensitive to effective inversion and storage temperature than to fill volumes. Conclusions Blood collection tubes with different RNA stabilizers present different robustness to common preanalytical variations.

Download Full-text

The stability of 25-hydroxyvitamin D in human blood during different sampling and storage conditions

Bone ◽

10.1016/j.bone.2009.03.206 ◽

2009 ◽

Vol 44 ◽

pp. S364

Author(s):

S. við Streym Thomsen⁎ ◽

L. Rejnmark ◽

P. Vestergaard ◽

L. Heickendorff ◽

L. Mosekilde

Keyword(s):

Human Blood ◽

Storage Conditions ◽

Hydroxyvitamin D ◽

25 Hydroxyvitamin D ◽

The Stability ◽

And Storage

Download Full-text

Special Collection and Storage Tubes for Blood Endotoxin and Cytokine Measurements

Clinical Chemistry ◽

10.1093/clinchem/38.5.764 ◽

1992 ◽

Vol 38 (5) ◽

pp. 764-765 ◽

Cited By ~ 25

Author(s):

H Redl ◽

S Bahrami ◽

G Leichtfried ◽

G Schlag

Keyword(s):

Tumor Necrosis Factor ◽

Blood Cells ◽

Tumor Necrosis ◽

Blood Collection ◽

Potential Source ◽

Special Collection ◽

And Storage ◽

Blood Collection Tubes ◽

Necrosis Factor

Abstract Commercially available blood-collection tubes may be contaminated with endotoxin (315 +/- 95 pg/tube) and could therefore be unsuitable for blood collection for endotoxin measurement. Plasma separation and storage are a potential source of contamination. To avoid contamination and error, we have developed new blood collection tubes that contain heparin free of endotoxin (LPS) and a gel to separate plasma and blood cells. The LPS content is less than 4 pg/tube. Samples can be stored and frozen without plasma withdrawal to preclude contamination. LPS recovery experiments have shown that the new blood-collection tubes do not bind LPS to the separation gel or vial wall. With these tubes, in vitro formation of tumor necrosis factor (404 +/- 163 ng/L in standard tubes vs less than 40 ng/L in special collection tubes) is minimized.

Download Full-text

Effects of storage temperature and time before centrifugation on ionized calcium in blood collected in plain vacutainer tubes and silicone-separator (SST) tubes.

Clinical Chemistry ◽

10.1093/clinchem/30.4.553 ◽

1984 ◽

Vol 30 (4) ◽

pp. 553-556 ◽

Cited By ~ 28

Author(s):

J Toffaletti ◽

N Blosser ◽

K Kirvan

Keyword(s):

Room Temperature ◽

Storage Temperature ◽

Ionized Calcium ◽

Blood Collection ◽

Healthy Individuals ◽

Blood Samples ◽

The Mean ◽

Whole Blood Samples ◽

The Stability ◽

Blood Collection Tubes

Abstract We studied the stability of ionized calcium and pH in samples stored at either room temperature or 4 degrees C, in centrifuged and uncentrifuged blood-collection tubes and in centrifuged tubes containing a silicone-separator gel (SST tubes). At room temperature, in uncentrifuged blood from healthy individuals, mean ionized calcium usually increased no more than 10 mumol/L per hour; at 4 degrees C it did not change detectably for 70 h. This stability was fortuitous, however: the concentrations of both hydrogen and lactate ions in these samples increased, apparently with offsetting effects on the concentration of ionized calcium. Blood stored for 70 h at 4 degrees C in centrifuged SST tubes, although showing a slightly greater change in ionized calcium, had less change of pH and no change in the ionized calcium corrected to pH 7.4. In 11 heparinized whole-blood samples from eight patients in intensive care, the mean change per hour in ionized calcium and pH after storage at room temperature was +10 mumol/L and -0.04 units, respectively.

Download Full-text

Vitamin D status and predictors of serum 25-hydroxyvitamin D concentrations in Western Australian adolescents

British Journal Of Nutrition ◽

10.1017/s000711451400186x ◽

2014 ◽

Vol 112 (7) ◽

pp. 1154-1162 ◽

Cited By ~ 14

Author(s):

Lucinda J. Black ◽

Sally A. Burrows ◽

Peter Jacoby ◽

Wendy H. Oddy ◽

Lawrence J. Beilin ◽

...

Keyword(s):

Vitamin D ◽

Mixed Model ◽

Linear Mixed Model ◽

Family Income ◽

Blood Collection ◽

Vitamin D Status ◽

International Consensus ◽

Hydroxyvitamin D ◽

25 Hydroxyvitamin D ◽

Western Australian

Despite the importance of skeletal growth during adolescence, there is limited research reporting vitamin D status and its predictors in adolescents. Using prospective data from the Western Australian Pregnancy Cohort (Raine) Study, we investigated vitamin D status and predictors of serum 25-hydroxyvitamin D (25(OH)D) concentrations in adolescents. Serum 25(OH)D concentrations were measured in the same participants at 14 and 17 years (n 1045 at both time points). The percentage of adolescents with serum 25(OH)D concentrations < 50, 50–74·9 and ≥ 75 nmol/l was reported year-round and by month of blood collection. We examined the predictors of serum 25(OH)D concentrations, including sex, race, month of blood collection, physical activity, BMI, family income, and Ca and vitamin D intakes (n 919 at 14 years; n 570 at 17 years), using a general linear mixed model. At 14 years, 31 % of adolescents had serum 25(OH)D concentrations between 50 and 74·9 nmol/l and a further 4 % had concentrations < 50 nmol/l. At 17 years, 40 % of adolescents had serum 25(OH)D concentrations between 50 and 74·9 nmol/l and 12 % had concentrations < 50 nmol/l. Caucasian ethnicity, being sampled at the end of summer, exercising more, having a lower BMI, a higher Ca intake and a higher family income were significantly associated with higher serum 25(OH)D concentrations. The proportion of adolescents with serum 25(OH)D concentrations < 50 nmol/l was low in this Western Australian cohort. There is a need for international consensus on defining adequate vitamin D status in order to determine whether strategies to increase vitamin D status in adolescents are warranted.

Download Full-text

Circulating 25-Hydroxyvitamin D Levels and Survival in Patients With Colorectal Cancer

Journal of Clinical Oncology ◽

10.1200/jco.2007.15.1027 ◽

2008 ◽

Vol 26 (18) ◽

pp. 2984-2991 ◽

Cited By ~ 205

Author(s):

Kimmie Ng ◽

Jeffrey A. Meyerhardt ◽

Kana Wu ◽

Diane Feskanich ◽

Bruce W. Hollis ◽

...

Keyword(s):

Colorectal Cancer ◽

Cancer Progression ◽

Proportional Hazards ◽

Cox Proportional Hazards ◽

Colorectal Carcinogenesis ◽

Health Study ◽

Blood Collection ◽

Hydroxyvitamin D ◽

25 Hydroxyvitamin D ◽

Overall Mortality

Purpose Higher plasma 25-hydroxyvitamin D3 (25(OH)D) levels are associated with a decreased incidence of colorectal cancer, but the influence of plasma 25(OH)D on the outcome of patients with established colorectal cancer is unknown. Patients and Methods We prospectively examined the association between prediagnosis 25(OH)D levels and mortality among 304 participants in the Nurses' Health Study (NHS) and the Health Professionals Follow-Up Study (HPFS) who were diagnosed with colorectal cancer from 1991 to 2002. Participants diagnosed within 2 years of blood collection were excluded. Patients were observed until death, June 2005 (NHS), or January 2005 (HPFS), whichever came first. The primary end point was overall mortality. Cox proportional hazards models were used to calculate hazard ratios (HR) adjusted for other risk factors for cancer survival. Results Higher plasma 25(OH)D levels were associated with a significant reduction in overall mortality (P for trend = .02). Compared with the lowest quartile, participants in the highest quartile had an adjusted HR of 0.52 (95% CI, 0.29 to 0.94) for overall mortality. A trend toward improved colorectal cancer–specific mortality was also seen (HR = 0.61; 95% CI, 0.31 to 1.19). The results remained unchanged after excluding patients diagnosed within 5 years of blood collection (P for trend = .04); the multivariate HR for overall mortality comparing extreme quartiles was 0.45 (95% CI, 0.19 to 1.09). Conclusion Among patients with colorectal cancer, higher prediagnosis plasma 25(OH)D levels were associated with a significant improvement in overall survival. Further study of the vitamin D pathway and its influence on colorectal carcinogenesis and cancer progression is warranted.

Download Full-text