scholarly journals Stability of parathyroid hormone ex vivo in haemodialysis patients

Author(s):  
T. K. Teal ◽  
M. Reed ◽  
P. E. Stevens ◽  
E. J. Lamb

Background: The stability of parathyroid hormone (PTH) in blood ex vivo is a significant practical problem for laboratories and clinicians. Several studies have suggested that PTH is more stable in blood collected into a potassium edetate (EDTA) preservative. Methods: To confirm that this was applicable to renal dialysis patients using our assay (Nichols chemiluminescence), we examined PTH stability in 13 patients with end-stage renal failure using three different blood collection tubes. Results: PTH remained stable in EDTA plasma for up to 48 h at room temperature. PTH was significantly reduced in serum collected into plain tubes after 2 h, and after 4 h in serum collected into serum separator tubes, at room temperature. Conclusion: In the assessment of renal osteodystrophy, the use of EDTA plasma can confer significant benefit, especially in busy laboratories where rapid frozen separation of blood may be hard to achieve.

Author(s):  
G E Levin ◽  
J A Nisbet

The stability of plasma parathyroid hormone-related protein (PTHrP) as measured by the Nichols Institute assay at room temperature was assessed over a period of 72 h in blood samples collected in protease inhibitor tubes and EDTA tubes at 0, 6, 24, 48 and 72 h from 10 patients with hypercalcaemia of malignancy. Mean plasma PTHrP concentrations in blood samples collected in protease inhibitor tubes remained stable for up to 48 h but had decreased by 10% at 72 h. The mean EDTA plasma PTHrP at zero time was 67% of the protease inhibitor tube value and this had fallen to 39% at 72 h. The stability of parathyroid hormone (PTH) in separated blood samples was also assessed by collection into heparin and plain tubes as well as EDTA and protease inhibitor tubes. Serum PTH concentrations progressively declined throughout the 72 h study period although the zero time values were significantly higher than corresponding plasma PTH concentrations. Plasma PTH concentrations appeared to be stable when blood was collected in heparin, EDTA and protease inhibitor tubes during the 72 h period, except in one subject with markedly elevated plasma amylase activity.


1984 ◽  
Vol 30 (4) ◽  
pp. 553-556 ◽  
Author(s):  
J Toffaletti ◽  
N Blosser ◽  
K Kirvan

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.


Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3207-3207
Author(s):  
Graham Smith ◽  
Loida Sheridan-Smith ◽  
Margaret A. Smith ◽  
Sam Pambakian ◽  
Jim Campbell

Abstract Platelet function is a vital factor in preventing bleeding irrespective of circulating platelet number. As a result, patients often have their surgery cancelled due to taking Aspirin or related medications. Similarly, patients bleed in the ITU setting when their counts and humoral coagulation are normal. This raises the concept of exhausted platelets which is often difficult to prove. The availability of a bedside technique which enumerates patients’ "functional" platelets, which is reliable and reproducible, represents a major step forward in determining whether these patients require platelet transfusion. Helena Laboratories, Beaumont, Texas, US have produced blood collection bottles which are coated with platelet agonists equine collagen (C), risocetin (R) or arachidonic acid (AA) in addition to 3.2mg sodium citrate. Our group is the first in Europe to use these bottles in association with point of care testing instrumentation (Horiba ABX Pentra, Chicksands, Beds, England). Samples were taken from normal, renal dialysis, cardiac catheterisation, haematological and ITU patients into K2EDTA and the above bottles and treated identically prior to counting. Samples taken into K2EDTA served as baseline controls whilst free platelets in the synchronously taken agonist samples represented non functional platelets (the normal functional ones having aggregated onto the inside wall of the tube). Data on 54 normal subjects showed that, when using C, only 6.2+/−3.1% of platelets were non functional (NF). Similar figures were found for R and AA. In dialysis patients with eGFR 4–25mls/min (n=32), the R NF% was 58.07+/−17.59%. In a similar group of renal patients on dialysis and Aspirin (n=12), the R NF% was 60.55+/−21.86% which was not statistically different from the non Aspirin group (p=0.955). In cardiac patients, all of whom were on Aspirin, C NF% was 35.23+/−23.12%, AA NF% 51.94+/−23.5% and R NF% 51.53+/−19.04%. C vs AA showed a significant p-value of 0.003and C vs R p=0.002. AA vs R was not significant at p=0.94. These data show that ristocetin is a surprisingly sensitive method of detecting non functional platelets in renal and Aspirin users. Classically, ristocetin is not thought of being a detector of Aspirin effect although data from Sloand et al. JASN (1997) 8(5),799 shows that ristocetin may detect renal effect on platelet GpIb-IX (CD42b). These data demonstrate the potential value of ristocetin aggregation ex vivo in determining the need for platelet transfusion and may provide clinicians with an improved and more logical rationale for such an expensive therapy.


2019 ◽  
Vol 37 (15_suppl) ◽  
pp. e14529-e14529
Author(s):  
Greg Sommer ◽  
Laura Fredriksen ◽  
Gabriella Iacovetti ◽  
Kyungjin Hong ◽  
Ulrich Schaff

e14529 Background: Sample quality is a critical consideration for high fidelity cell-free DNA (cfDNA) testing. Oncological cfDNA tests used for liquid biopsy typically employ specialty blood collection tubes containing chemical preservatives to minimize degradation of samples prior to lab testing. Here we describe a newly developed device, Zero Delay Plasma– a portable centrifuge and disc system designed to immediately isolate cell-free plasma at the point of blood draw – and evaluate its performance against the Streck cfDNA collection tube. Methods: Whole blood was collected, processed, and stored at room temperature for up to 7 days with both the Zero Delay Plasma system and the Streck cfDNA blood collection tube. Sample hemolysis was measured via cell-free hemoglobin. Genomic contamination and cfDNA signal-to-noise were evaluated by qPCR and electrophoresis, comparing signal from target 150-200bp cfDNA to contaminating longer length genomic sequences in the sample. 2 sets of hemolysis experiments, 2 sets of electrophoresis experiments and 4 sets of qPCR experiments were conducted. Results: Plasma processed with the Zero Delay Plasma system yielded ~4X lower hemolysis levels, ~10X lower genomic contamination, and ~20X higher cfDNA signal-to-noise compared to the Streck cfDNA collection tube after 7 days of storage at room temperature. Conclusions: The Zero Delay Plasma system minimizes sample degradation and analytical background signal for cfDNA testing by immediately removing cells and other contaminants at the point of blood collection. Clinical evaluations are in process.


2004 ◽  
Vol 50 (9) ◽  
pp. 1713-1714 ◽  
Author(s):  
Teresa K Teal ◽  
Joanna L Wood ◽  
Paul E Stevens ◽  
Edmund J Lamb

2003 ◽  
Vol 49 (12) ◽  
pp. 2012-2019 ◽  
Author(s):  
Sue Xiang ◽  
Rachel Denver ◽  
Michael Bailey ◽  
Henry Krum

Abstract Background: Salivary endothelin (ET) concentrations have been shown to correlate with disease severity in patients with chronic heart failure (CHF). We undertook the present study to evaluate the stability of salivary ET under different handling conditions to assess its suitability as a biochemical marker in screening, diagnosis, and management of CHF. Methods: Saliva samples were collected from healthy individuals and/or CHF patients, subjected to different handling conditions, and then stored at −80 °C until assayed by an ELISA for ET. Results: Salivary ET concentrations showed a time-dependent increase during storage at room temperature. After 72 h of incubation at room temperature, ET increased ∼2.8-fold (P = 0.03). Simultaneously, salivary big ET showed a time-dependent 11.2-fold decrease (P <0.0001). This activity was blocked by an ET-converting enzyme (ECE) inhibitor, suggesting that these changes were attributable to ECE-dependent cleavage of endogenous big ET in saliva. Ex vivo conversion was also observed when samples were stored at 4 °C, but the magnitude of these changes was markedly smaller (P <0.0001). Posture also affected salivary ET concentrations in CHF patients. With a change from supine to seated rest, salivary ET concentrations increased 1.5- and 1.8-fold after 20 and 40 min, respectively (P = 0.01). With a return to supine rest, salivary ET concentrations returned to baseline concentrations (P = 0.008). Conclusions: These data suggest that saliva sampling and handling conditions could markedly affect measurement of salivary ET. In particular, care should be taken to minimize ECE-dependent enzymatic conversion of endogenous big ET in saliva.


2018 ◽  
Vol 56 (9) ◽  
pp. 1476-1482
Author(s):  
Andre Valcour ◽  
Claudia Zierold ◽  
Frank A. Blocki ◽  
Douglas M. Hawkins ◽  
Kevin J. Martin ◽  
...  

Abstract Background: Over the past few decades, parathyroid hormone (PTH) immunoassays have progressed through successive generations resulting in increased specificity and accuracy for detecting circulating PTH. With the introduction of third-generation assays, in which the biologically active PTH(1-84) is specifically targeted, the PTH(7-84) and other fragments are not detected. The specific recognition of only PTH(1-84) whole molecule allows for more reliable standardization and calibration than with the existing assays. Methods: Samples from patients on hemodialysis or with primary hyperparathyroidism and apparently healthy subjects were examined in different collection matrices (EDTA plasma, unspun EDTA plasma and SST) stored for 0, 24 or 72 h at room temperature to reflect the prevailing sample collection methods, shipping and processing conditions of centralized labs in the United States. Samples were analyzed by the LIAISON 1-84 PTH and N-TACT assays, and by three additional commercially available intact PTH assays. Results: Defined samples, prepared using two different standards (WHO 95/646 international standard and the synthetic Bachem PTH(1-84)), show little bias with the LIAISON 1-84 PTH assay, but not with the other intact PTH assays. Furthermore, PTH is stable for up to 72 h in plasma, but less stable in serum beyond 24 h. Conclusions: The FDA-approved LIAISON 1-84 PTH assay is accurate and reliably measures the biologically active PTH molecule in plasma or serum stored at room temperature for up 72 and 24 h, respectively.


2001 ◽  
Vol 100 (6) ◽  
pp. 609-611 ◽  
Author(s):  
Matthew L. P. HOWSE ◽  
Maureen LEONARD ◽  
Michael VENNING ◽  
Laurence SOLOMAN

Metabolic acidosis frequently complicates end-stage renal failure. In haemodialysis patients its severity is usually monitored by measurement of the total CO2 (TCO2) level. Samples from ‘satellite dialysis’ patients are often stored prior to analysis. We investigated the affect of storage of 21 samples for 24 h under different conditions prior to analysis. If samples were stored at room temperature the TCO2 fell from 22.7±4.2 mmol/l to 21.6±3.7 mmol/l (P = 0.001). If the same samples were spun and stored at 4 °C the TCO2 was 22.4±3.9 mmol/l (P = not significant). We conclude that the magnitude in the fall of TCO2 stored at room temperature for 24 h is unlikely to be clinically significant and can be prevented by spinning the sample and refrigerating it.


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