Applying Best Practice to Converting Blood Collection Tube Suppliers: Overcoming Laboratory Supply Shortages

Introduction/Objective Changing blood tube suppliers is a complex process that requires systematic validation. It can serve to expand a laboratory’s options during a supply shortage, and can lead to cost savings. Just prior to the pandemic, our laboratory underwent a large-scale conversion of blood tube suppliers after successful validation of serum, plasma, and whole blood tubes for blood bank, chemistry, immunology, hematology, coagulation, molecular diagnostics, and flow cytometry. Methods/Case Report First, we designed a patient consent form for collecting extra blood samples. Per CLSI standards, validation of blood tubes is needed for each testing methodology but not for each analyte. Hence, we selected high-impact assays to represent each testing methodology used by our instrument platforms. We designed comparison studies to test the new blood tubes for result accuracy, precision and specimen stability that covered an assay’s reportable range. Allowable error limits were set based on Westgard and CAP guidelines. Spiked specimens were used when positive patient samples were not feasible. We also took the opportunity to optimize the blood tube sizes, e.g., converting the lavender-top tubes from 2 ml (vendor A) to 3 ml (vendor B). We then confirmed with our reference labs the acceptability of the new vendor’s products, and administered an electronic survey to solicit staff feedback. Finally, we coordinated supply chain, formulary changes and test catalog updates. Results (if a Case Study enter NA) Data analyses showed 100% acceptable performance of the new supplier’s blood tubes. The survey showed that our phlebotomists supported the new products, citing their ease of use and good vacuum. Our hematology technologists provided favorable feedback on the larger lavender-top tubes that reduced the number of insufficient samples. On the economic front, this supplier conversion has yielded a 26% cost savings. Conclusion To our knowledge, our study is the most comprehensive of all blood collection device comparison analyses published. Our validation strategies that were designed to comply with best practice standards have led to our confidence in the interchangeability of the new and old blood tube products. This initiative serves to elucidate a protocol to add vendor options that can be replicated by other laboratories to mitigate blood tube supply shortages and backorders. It has helped us control supply costs without compromising quality.

The effect of pre-analytical and physiological variables on cell-free DNA fragmentation.

Assays that account for the biological properties and fragmentation of cell-free DNA (cfDNA) can improve the performance of liquid biopsy. However, pre-analytic and physiological differences between individuals on fragmentomic analysis are poorly defined. We analyzed the impact of collection tube, plasma processing time and physiology on the size distribution of cfDNA, their genome-wide representation and sequence diversity at the cfDNA fragment-ends using shallow Whole Genome Sequencing. We observed that using different stabilizing collection tubes, or processing times does not affect the cfDNA fragment sizes, but can impact the genome-wide fragmentation patterns and fragment-end sequences of cfDNA. In addition, beyond differences depending on the gender, the physiological conditions tested between 63 individuals (age, body mass index, use of medication and chronic conditions) minimally influenced the outcome of fragmentomic methods. Our results highlight that fragmentomic approaches have potential for implementation in the clinic, pending clear traceability of analytical and physiological factors.

A Comparison of Serum and Plasma Blood Collection Tubes for the Integration of Epidemiological and Metabolomics Data

Blood is a rich biological sample routinely collected in clinical and epidemiological studies. With advancements in high throughput -omics technology, such as metabolomics, epidemiology can now delve more deeply and comprehensively into biological mechanisms involved in the etiology of diseases. However, the impact of the blood collection tube matrix of samples collected needs to be carefully considered to obtain meaningful biological interpretations and understand how the metabolite signatures are affected by different tube types. In the present study, we investigated whether the metabolic profile of blood collected as serum differed from samples collected as ACD plasma, citrate plasma, EDTA plasma, fluoride plasma, or heparin plasma. We identified and quantified 50 metabolites present in all samples utilizing nuclear magnetic resonance (NMR) spectroscopy. The heparin plasma tubes performed the closest to serum, with only three metabolites showing significant differences, followed by EDTA which significantly differed for five metabolites, and fluoride tubes which differed in eleven of the fifty metabolites. Most of these metabolite differences were due to higher levels of amino acids in serum compared to heparin plasma, EDTA plasma, and fluoride plasma. In contrast, metabolite measurements from ACD and citrate plasma differed significantly for approximately half of the metabolites assessed. These metabolite differences in ACD and citrate plasma were largely due to significant interfering peaks from the anticoagulants themselves. Blood is one of the most banked samples and thus mining and comparing samples between studies requires understanding how the metabolite signature is affected by the different media and different tube types.

Microfluidic Fabrication of Helical Ca-Alginate Hydrogel Fibers

Helix is a sophisticated structure in nature and has many unique functions which makes it possible to store more information and energy, even receive more sensitive signals. Besides, as an effective method for preparing hydrogel fibers, microfluidic spinning has achieved unprecedented development in the past decade. However, hydrogel fiber with helical structure has began to be studied only in recent years. In this paper, the helical hydrogel fibers were prepared by the microfluidic spinning method. The microfluidic chip was assembled by PDMS connector, collection tube, inner and outer channels. Sodium alginate (SA) and calcium chloride were used as the core fluid and sheath fluid, respectively. By designing and adjusting the length of the chip, changing the concentration of SA and the ratio of two flow rates (inner flow rate/outer flow rate), a continuous and uniform helical hydrogel fiber was prepared. The relationships between the diameter of the fiber, the pitch of the helix and the concentration of SA, the ratio of two flow rates were discussed. The results showed that the diameter of the fiber was mainly affected by the core fluid. Within a certain range, as the concentration of SA increased, the diameter of the fiber increased. Besides, the pitch of the helix was greatly affected by the flow rate of sheath fluid. As the velocity of the sheath fluid increased, the pitch of the fiber increased. Such helical fiber could be used in micro sensors when added some conductive materials or crosslinked with some temperature responsive polymers such as N-isopropylacrylamide.

Fabrication of Multi-Layered Microspheres Based on Phase Separation for Drug Delivery

In this work, we used a co-flow microfluidic device with an injection and a collection tube to generate droplets with different layers due to phase separation. The phase separation system consisted of poly(ethylene glycol) diacrylate 700 (PEGDA 700), PEGDA 250, and sodium alginate aqueous solution. When the mixture droplets formed in the outer phase, PEGDA 700 in the droplets would transfer into the outer aqueous solution, while PEGDA 250 still stayed in the initial droplet, breaking the miscibility equilibrium of the mixture and triggering the phase separation. As the phase separation proceeded, new cores emerged in the droplets, gradually forming the second and third layers. Emulsion droplets with different layers were polymerized under ultraviolet (UV) irradiation at different stages of phase separation to obtain microspheres. Microspheres with different layers showed various release behaviors in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF). The release rate decreased with the increase in the number of layers, which showed a potential application in sustained drug release.

EXPRESS: Evaluation of the quick-clotting serum separator tube, VQ-TubeTM, for clinical chemistry and thyroid hormone assays

Background: The type of blood collection tube affects specimen quality and laboratory results. Because plasma specimens have a shorter processing time compared to serum specimens, emergency biochemistry tests use plasma. However, serum specimens remain stable after centrifugation and show more accurate results than plasma. Therefore, a quick-clotting serum separator tube (SST) is expected to be useful for shorter turnaround times and accurate results. We evaluated a new quick-clotting SST VQ-Tube^TM (AB Medical, Korea) for clinical chemistry and thyroid hormone assays. Methods: One hundred volunteers from four university hospitals were recruited and peripheral blood samples were collected in quick-clotting SST VQ-Tubes^TM and the commonly used SST V-Tubes^TM. The obtained specimens were used for 16 clinical chemistry assays and three thyroid hormone assays. Results: The differences (%) in the test results obtained from the samples in each tube satisfied the allowable difference ranges (19 assays). The differences in the test results between the tubes satisfied the desired specifications for accuracy except for the glucose results (2.75%). The paired t-test revealed significant differences between the results of six assays but each set of results showed a good correlation. Samples were visually inspected for serum clarity and gel barrier integrity, and incomplete clotting reactions and hemolyzed serum were not observed. Conclusion: The new quick-clotting VQ-Tube^TM demonstrated reliable test results compared to the commonly used SST V-Tube^TM. This quick-clotting tube will provide fast test results with adequately separated serum specimens, especially for patients who need fast tests.

Comparative metabolomics studies of blood collected in streck and heparin tubes from lung cancer patients

Metabolomics analysis of blood from patients (n = 42) undergoing surgery for suspected lung cancer was performed in this study. Venous and arterial blood was collected in both Streck and Heparin tubes. A total of 96 metabolites were detected, affected by sex (n = 56), collection tube (n = 33), and blood location (n = 8). These metabolites belonged to a wide array of compound classes including lipids, acids, pharmaceutical agents, signalling molecules, vitamins, among others. Phospholipids and carboxylic acids accounted for 28% of all detected compounds. Out of the 33 compounds significantly affected by collection tube, 18 compounds were higher in the Streck tubes, including allantoin and ketoleucine, and 15 were higher in the Heparin tubes, including LysoPC(P-16:0), PS 40:6, and chenodeoxycholic acid glycine conjugate. Based on our results, it is recommended that replicate blood samples from each patient should be collected in different types of blood collection tubes for a broader range of the metabolome. Several metabolites were found at higher concentrations in cancer patients such as lactic acid in Squamous Cell Carcinoma, and lysoPCs in Adenocarcinoma and Acinar Cell Carcinoma, which may be used to detect early onset and/or to monitor the progress of the cancer patients.

Cloning, Characterization and Bioinformatics Analysis of the Sequences of miR-10a and miR-10b in Sheep (Hu sheep)

Background: MicroRNAs (miRNAs) are active regulators of numerous biological and physiological processes and play an important role in the regulation of animal ovaries and other reproductive related organs. To understand the molecular mechanisms of miR-10 family, we investigated the molecular characteristics and the relative expression of sheep miR-10a and miR-10b (miR-10a/b) and conducted bioinformatics analysis.Methods: During the period 2018-2019 total of 20 samples including blood and tissues such as hypothalamus, pituitary and ovary were collected from the Hu sheep raised in the National Meat Sheep Experimental Station of China (Luoyang City, Henan Province, China). Blood was collected from jugular vein by vacuum and anticoagulation blood collection tube and stored in refrigerator at -20°C. The tissues were placed into the cryopreservation tube treated with Diethy pyrocarbonate (DEPC) water and stored in liquid nitrogen. All the samples were processed for isolation and confirmed with biochemical analysis and Polymerase chain reaction (PCR) and Real-time fluorescence quantitative PCR.Result: The target genes were predicted by three kinds of target gene predicting software. The function of target genes and their involved pathways were obtained by gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The relative expression of miR-10a/b in sheep ovary was extremely significantly higher than that in hypothalamus and pituitary gland (P less than 0.01). The relative expression of miR-10b in ovary or pituitary was extremely significantly higher than that of miR-10a (P less than 0.01) and the relative expression of miR-10b in hypothalamus was significantly higher than that of miR-10a (P less than 0.05). These results serve as a foundation for further study on the Sheep miR-10 family.

Brush Licking Toxicity: Toxin & Microorganism Growth On Paint Brushes

Introduction/Objective A common practice utilized by painters is licking their paint brush bristles to form a sharp point for fine lines and details. Literature details artists careless around their mediums have a higher incidence of becoming ill, yet if their brushes harbor any toxic substances or pathogenic bacteria it is currently unknown. Therefore, this study aims to determine if there is risk associated with brush licking, by determining if pathogenic bacteria and/or heavy metals are present on the brushes of volunteer artists. Methods We obtained 17 volunteer paint brushes and inoculated specialized collection and transport media (E- Swab, BD Biosciences, San Jose, CA) while the volunteer completed a qualitative de-identified survey indicating brush licking status. Brushes were swirled in the E-swab collection tube, then subbed to sheep blood agar plates and chocolate agar plates for bacteria growth analysis. Matrix-assisted laser desorption/ionization time-of-flight (MALDI- TOF) technology was used for identification. With the remaining E-Swab media, lead (3M, St. Paul, MN) and arsenic (HACH, Loveland, CO) testing using commercial kits was done. MALDI-TOF identification and heavy metal testing results were then compared to survey results. Results One of 17 specimens (6%) had pathogenic bacteria identified (Pseudomonas aeruginosa). This specimen’s survey also indicated routine brush licking. The sensitivity and specificity of MALDI-TOF for Pseudomonas aeruginosa is 96.67% and 97.87%, respectively. The remaining 16/17 (94%) had normal flora present. Importantly, the specimen containing Pseudomonas aeruginosa also had high levels of arsenic at 50 ppb. None of the specimen tested contained lead. Conclusion Our results indicated there is indeed some risk associated with brush licking. However, due to the small sample size, statistical significance could not be determined. Nonetheless, with the lack of knowledge surrounding this subject, it is beneficial to further explore and educate painters on the toxicities of brush licking.