Preanalytical robustness of blood collection tubes with RNA stabilizers

2019 ◽  
Vol 57 (10) ◽  
pp. 1522-1529 ◽  
Author(s):  
Chiara Stellino ◽  
Gaël Hamot ◽  
Camille Bellora ◽  
Johanna Trouet ◽  
Fay Betsou
Keyword(s):  
Storage Temperature ◽  
Regulation Of Gene Expression ◽  
Analysis Data ◽  
Blood Collection ◽  
Becton Dickinson ◽  
Rna Integrity ◽  
Qrt Pcr ◽  
Rna Integrity Number ◽  
Significant Difference ◽  
Blood Collection Tubes

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.

Residual Negative Pressure in Vacuum Blood-Collection Tube and Hemolysis in Pediatric Blood Specimens

2019 ◽  
Vol 51 (1) ◽  
pp. 41-46
Author(s):  
Jing Hu ◽  
Qiao-Xin Zhang ◽  
Tong-Tong Xiao ◽  
Mei-Chen Pan ◽  
Ying-Mu Cai
Keyword(s):  
Negative Pressure ◽  
Blood Collection ◽  
Free Hemoglobin ◽  
Significant Difference ◽  
Collection Tube ◽  
Biochemical Analyzer ◽  
Blood Specimens ◽  
Adult Volunteers ◽  
Blood Collection Tubes ◽  
Blood Collection Tube

ABSTRACT Objective To determine a method to reduce specimen hemolysis rates in pediatric blood specimens. Methods A total of 290 blood specimens from pediatric patients were classified into the capped group or uncapped group. The hemolysis index and levels of lactate dehydrogenase (LDH) were measured using an automated biochemical analyzer. Also, we performed a paired test to measure the concentration of free hemoglobin in specimens from 25 randomly selected healthy adult volunteers, using a direct spectrophotometric technique. Results The hemolytic rate of capped specimens was 2-fold higher than that of uncapped specimens. We found significant differences for LDH. Also, there was a significant difference in the concentration of free hemoglobin in the random-volunteers test. Conclusions Eliminating the residual negative pressure of vacuum blood-collection tubes was effective at reducing the macrohemolysis and/or microhemolysis rate.

scholarly journals The kinetics of ageing in dry-stored seeds: a comparison of viability loss and RNA degradation in unique legacy seed collections

2018 ◽  
Vol 123 (7) ◽  
pp. 1133-1146 ◽  
Author(s):  
Margaret B Fleming ◽  
Lisa M Hill ◽  
Christina Walters
Keyword(s):  
Storage Time ◽  
Storage Temperature ◽  
Rna Degradation ◽  
Seed Longevity ◽  
Rna Integrity ◽  
Viability Loss ◽  
Rna Integrity Number ◽  
Seed Ageing ◽  
Mode Of Detection ◽  
Ageing Rate

Abstract Background and Aims Determining seed longevity by identifying chemical changes that precede, and may be linked to, seed mortality, is an important but difficult task. The standard assessment, germination proportion, reveals seed longevity by showing that germination proportion declines, but cannot be used to predict when germination will be significantly compromised. Assessment of molecular integrity, such as RNA integrity, may be more informative about changes in seed health that precede viability loss, and has been shown to be useful in soybean. Methods A collection of seeds stored at 5 °C and 35–50 % relative humidity for 1–30 years was used to test how germination proportion and RNA integrity are affected by storage time. Similarly, a collection of seeds stored at temperatures from −12 to +32 °C for 59 years was used to manipulate ageing rate. RNA integrity was calculated using total RNA extracted from one to five seeds per sample, analysed on an Agilent Bioanalyzer. Results Decreased RNA integrity was usually observed before viability loss. Correlation of RNA integrity with storage time or storage temperature was negative and significant for most species tested. Exceptions were watermelon, for which germination proportion and storage time were poorly correlated, and tomato, which showed electropherogram anomalies that affected RNA integrity number calculation. Temperature dependencies of ageing reactions were not significantly different across species or mode of detection. The overall correlation between germination proportion and RNA integrity, across all experiments, was positive and significant. Conclusions Changes in RNA integrity when ageing is asymptomatic can be used to predict onset of viability decline. RNA integrity appears to be a metric of seed ageing that is broadly applicable across species. Time and molecular mobility of the substrate affect both the progress of seed ageing and loss of RNA integrity.

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

PLoS ONE ◽  
2016 ◽  
Vol 11 (11) ◽  
pp. e0166327
Author(s):  
Songlin Yu ◽  
Weiyan Zhou ◽  
Xinqi Cheng ◽  
Huiling Fang ◽  
Ruiping Zhang ◽  
...  
Keyword(s):  
Storage Temperature ◽  
Blood Collection ◽  
Hydroxyvitamin D ◽  
25 Hydroxyvitamin D ◽  
And Storage ◽  
Blood Collection Tubes

scholarly journals Immunoassay Interference by a Commonly Used Blood Collection Tube Additive, the Organosilicone Surfactant Silwet L-720

2005 ◽  
Vol 51 (10) ◽  
pp. 1874-1882 ◽  
Author(s):  
Raffick AR Bowen ◽  
Yung Chan ◽  
Mark E Ruddel ◽  
Glen L Hortin ◽  
Gyorgy Csako ◽  
...  
Keyword(s):  
Solid Phase ◽  
Blood Collection ◽  
Becton Dickinson ◽  
Total Triiodothyronine ◽  
Collection Tube ◽  
Capture Antibody ◽  
Blood Collection Tubes ◽  
Blood Collection Tube ◽  
Selection Of ◽  
Serum Components

Abstract Background: A small number of immunoassays on several different types of analyzers were recently adversely affected by tube additives in Becton Dickinson (BD) Vacutainer® SST™, SST II, and Microtainer™ blood collection tubes. We examined the effect of a commonly used tube surfactant, Silwet™ L-720, on immunoassays and the mechanism for the interference. Methods: Immunoassays were performed on serum supplemented with Silwet L-720 on the IMMULITE™ 2500 and AxSYM™ analyzers. Direct effects of the surfactant on the chemiluminescent detection step of immunoassays and on antibody immobilization on the solid phase were examined. Results: Increasing the final surfactant concentration from 0 to 400 mg/L in serum significantly increased (∼51%) the apparent total triiodothyronine (TT3) concentrations measured on the IMMULITE 2500 but not the AxSYM analyzer. Several other competitive, but not noncompetitive, assays were also significantly affected by the surfactant on the IMMULITE 2500 analyzer. The effect was independent of serum components, and the surfactant had no direct effect on chemiluminescence reactions. The capture antibody, however, was displaced from the solid phase by incubation with solutions containing surfactant under conditions similar to the IMMULITE TT3 assay. Conclusions: The Silwet L-720 surfactant, which is used to coat the inner surfaces of tubes, appears to account for previously reported immunoassay interference by BD Vacutainer SST blood collection tubes. One of the mechanisms for the interference is the desorption of antibodies from the solid phase by the surfactant. The results identify an important factor in the selection of suitable blood collection tube surfactants and provide an approach for solving similar tube-assay interference problems in the future.

scholarly journals homeRNA: A self-sampling kit for the collection of peripheral blood and stabilization of RNA

2021 ◽  
Author(s):  
Amanda J. Haack ◽  
Fang Yun Lim ◽  
Dakota S. Kennedy ◽  
John H. Day ◽  
Karen N. Adams ◽  
...  
Keyword(s):  
Whole Blood ◽  
Peripheral Blood ◽  
Rna Degradation ◽  
Blood Collection ◽  
Rna Integrity ◽  
Rna Integrity Number ◽  
Collection Device ◽  
Location Parameters ◽  
Rna Stabilization ◽  
Gene Panels

ABSTRACTGene expression analysis (e.g., targeted small gene panels, transcriptomics) from whole blood can elucidate mechanisms of immune function and aid in the discovery of biomarkers. Conventional in-clinic venipuncture offers only a small snapshot of our broad immune landscape as immune responses may occur outside of the time and location parameters available for conventional venipuncture. A self-operated method that enables flexible sampling of liquid whole blood coupled with an immediate stabilization of cellular RNA is instrumental in facilitating capture and preservation of acute or transient immune fluxes. To this end, we developed homeRNA: a kit that allows for self-collection of peripheral blood (∼0.5 mL) and immediate stabilization of cellular RNA, using the Tasso-SST™ blood collection device paired with a specially designed stabilizer tube containing RNAlater™. To assess the usability and feasibility of homeRNA for self-collection and stabilization of whole blood RNA, we conducted a pilot study (n = 41 participants) where we sent homeRNA to participants aged 21-69, located across 10 US states (94% successful blood collections, n = 51). Among participants who successfully collected blood, 91% reported no or minimal pain/discomfort using the kit (n = 35), and 77% reported easy or somewhat easy stabilization protocol. Total RNA yield from the stabilized samples ranged between 0.24 µg and 5.99 µg (mean = 1.65 µg), while RNA Integrity Number (RIN) values were above 7.0 (mean = 7.9), indicating limited RNA degradation. Results from this study demonstrate the self-collection and RNA stabilization of whole blood with homeRNA by participants themselves, in their own home.

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

1984 ◽  
Vol 30 (4) ◽  
pp. 553-556 ◽  
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.

Comparison of some biochemical tests in different blood collection tubes in hemodialysis patients

2019 ◽  
Vol 45 (1) ◽  
pp. 26-36 ◽  
Author(s):  
Arzu Kösem ◽  
Canan Topçuoğlu ◽  
Sevilay Sezer ◽  
Şimal Köksal Cevher ◽  
Ezgi Coşkun Yenigün ◽  
...  
Keyword(s):  
Clinical Chemistry ◽  
Statistical Significance ◽  
Blood Collection ◽  
Test Results ◽  
Biochemical Tests ◽  
Becton Dickinson ◽  
Two Samples ◽  
Clinically Significant ◽  
Roche Diagnostics ◽  
Blood Collection Tubes

Abstract Objective Blood collection tubes (BCTs) related interferences in test results can adversely influence on patient outcomes. We compared test results of samples in BD (Becton-Dickinson, Franklin Lakes, NJ, USA) Vacutainer Serum Separator Tubes (SST), BD Vacutainer® Barricor™ Plasma BCTs (Barricor™) and BD Vacutainer® Rapid Serum Tube (RST). Materials and methods Thirty-two samples were obtained from patients after the hemodialysis were included in this study. Eight routine clinical chemistry parameters (AST, creatinin, urea, PTH, glucose, LDH, K, calcium) were measured on Roche Cobas Analyzer (Roche Diagnostics, North America). The results of samples obtained from RST and Barricor™ were compared with SST as reference tubes. Results Results of Glucose, K, Urea, PTH from the SST and Barricor™ were statistically significantly different (p = 0.017, p < 0.001, p = 0.011, p < 0.001, respectively). In addition, results of PTH, LDH from SST and RST were significantly different (p < 0.001, p = 0.019). However, statistical significance of test results was not clinically significant for the biochemical parameters. Conclusion Working with Barricor™ may provide not just a fast, clean, high-quality plasma samples, safety results, but also time and cost-effectivity. Therefore, these types of tubes, which are less costly than other BCTs, may be preferred to obtain plasma.

Suitability of Becton Dickinson Vacutainer rapid serum tube for collecting and storing blood samples for antibiotic and anticonvulsant drug monitoring

2014 ◽  
Vol 67 (9) ◽  
pp. 807-810 ◽  
Author(s):  
Ronald Yan ◽  
David Colantonio ◽  
Pui-Yuen Wong ◽  
Yu Chen
Keyword(s):  
Drug Monitoring ◽  
Whole Blood ◽  
Drug Stability ◽  
Anticonvulsant Drug ◽  
Therapeutic Drug ◽  
Blood Collection ◽  
Becton Dickinson ◽  
Drug Concentrations ◽  
Plain Tube ◽  
Blood Collection Tubes

AimsTo investigate the suitability of newly developed Becton Dickinson Vacutainer rapid serum tube (RST) for therapeutic drug monitoring of antibiotics and anticonvulsants.MethodsTwo pools of citrated whole blood were created by spiking high and low concentrations of gentamicin, vancomycin, phenytoin, lamotrigine and carbamazepine. After recalcification with 15 mmol/L calcium chloride, spiked whole blood was added into four different Becton Dickinson blood collection tubes: RST, serum separator tube, red top tube and polyethylene plain tube. Serum aliquots were collected at baseline (0 h), 2 h, 24 h, day 3 and day 7. Drug concentrations were measured in batch by HPLC and the Architect c8000.ResultsGentamicin and vancomycin concentrations were stable up to 7 days in all 4 blood collection tubes. Anticonvulsants results for the RST were stable and did not deviate substantially from those of the red top and plain tubes, and demonstrated better performance than the serum separator tubes that showed significant (≥10% bias, p<0.05) decrease in phenytoin and carbamazepine levels after 3 days of storage.ConclusionsThe RST provides acceptable drug stability over the course of 7 days for gentamicin, vancomycin, phenytoin and lamotrigine and over 3 days for carbamazepine.

scholarly journals Effect of Blood Collection Tubes on Total Triiodothyronine and Other Laboratory Assays

2005 ◽  
Vol 51 (2) ◽  
pp. 424-433 ◽  
Author(s):  
Raffick AR Bowen ◽  
Yung Chan ◽  
Joshua Cohen ◽  
Nadja N Rehak ◽  
Glen L Hortin ◽  
...  
Keyword(s):  
Clinical Chemistry ◽  
Reference Interval ◽  
Blood Collection ◽  
Serum Samples ◽  
Becton Dickinson ◽  
Total Triiodothyronine ◽  
Patient Values ◽  
Clinically Significant ◽  
Age Range ◽  
Blood Collection Tubes

Abstract Background: Increased total triiodothyronine (TT3) assay results in apparently euthyroid patients triggered an investigation of the effect of blood collection tubes on serum TT3 and other laboratory assays. Methods: We examined potential assay interference for three types of tubes: plastic Greiner Bio-One™ Vacuette™; glass Becton Dickinson (BD) Vacutainer™; and plastic BD Vacutainer SST™ tubes. Serum samples from apparently healthy volunteers (age range, 30–60 years; 15 males and 34 females) were collected in different tube types and analyzed in 17 immunoassays (n = 49), 30 clinical chemistry tests (n = 20), and 33 immunology assays (n = 15). Tube effects were also examined by adding pooled serum to different tube types. Results: TT3 values, when measured by the IMMULITE™ 2000 but not the AxSYM™ analyzer, were significantly higher (P &lt;0.0001) for SST (2.81 nmol/L) than either glass (2.15 nmol/L) or Vacuette (2.24 nmol/L) tubes. The effect was large enough to substantially shift the distribution of patient values, increasing the percentage of values above the reference interval from 11.3% to 35.8%. The degree of interference from SST tubes on TT3 differed among various tube lots and could be attributed to a tube additive shared by other plastic tubes. Results from several other tests statistically differed among tube types, but differences were not considered to be clinically significant. Conclusions: Assay interferences from blood collection tubes represent challenges to clinical laboratories because they are not detected by the usual quality-control or proficiency testing programs. Laboratories can, however, address this problem by monitoring distribution of patients’ results.

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