Clinical evaluation of lysis-centrifugation technique and a biphasic bottle system for blood culture

The lysis centrifugation technique (Isolator, DuPont) for blood culture was compared with a system with biphasic medium in bottles. The Isolator was filled with 10 ml of blood once. One aerobic and one anaerobic bottle were injected 3 times with 2.5 ml of blood each. Organisms were detected in 90/748 blood cultures; 26 of which were contaminants. 34 pathogens were detected by both methods, 12 with the Isolator only and 18 with the 3 bottle pairs only. The first pair of bottles revealed 45/52 isolates, the second and third pairs gave an additional 6 and 1 isolate respectively. The contamination rate was 2.3% for the Isolator, which is lower than earlier reported, and 1.3% for the bottles. The most common pathogens were Escherichia coli, Pseudomonas aeruginosa and Streptococcus pneumoniae. The Isolator gave a faster diagnosis in 23 of the 34 cases. No decrease in the recovery rate was seen after 8 h, the longest recommended transport time for the Isolator tubes. One Isolator gave the same yield as the first pair of bottles and combining the methods increased the yield 25% compared to either method alone.     

Pilot study of self-measurement of blood glucose using the dextrostix-eyetone system for juvenile-onset diabetes

Summary Self-measurement of blood glucose was performed by eight insulin-dependent juvenile-onset diabetics. The patients were well motivated and improved control of diabetes was obtained. The method would seem to be useful for juvenile-onset diabetics, especially of the unstable type and for pregnant diabetics.     

Clotting time analysis of citrated blood samples is strongly affected by the tube used for blood sampling.

Our aim was to establish whether differences in clotting times for recalcified blood and plasma samples might be explained by the use of different blood collection tubes. Samples obtained from different plastic vacuum tubes were recalcified and clotting times determined by free oscillation rheometry. The clotting times for blood collected in Vacutainer (Becton Dickinson, Rutherford, New Jersey, USA) or Vacuette (Greiner Bio-One, Kremsmünster, Austria) tubes decreased with time, with maximal effect after 30 min. Blood from Monovette (Sarstedt, Nümbrecht, Germany) tubes displayed longer clotting times, which did not decrease with time. Clotting times for plasma prepared after 1 h storage in Vacutainer or Vacuette tubes were unaffected by subsequent addition of corn trypsin inhibitor to inhibit factor XIIa, although an antibody against factor XI prolonged the clotting time markedly. In Monovette plasma, both corn trypsin inhibitor and anti-factor XI effectively prolonged the clotting time. When corn trypsin inhibitor or anti-factor XI was added to the tubes before blood collection, but both additions clearly prolonged the clotting times in all types of tubes, even though corn trypsin inhibitor was less effective in whole blood. Antibodies against human tissue factor did not affect the clotting times. The amounts of platelet or leukocyte microparticles in plasma were low and similar in all tubes. This indicates that blood collection in Vacutainer or Vacuette tubes induces a rapid activation of factor XII and factor XI.     

Pilot study of oxygen transport rate of banked red blood cells

Background and Objectives  Dynamic oximetry provides a new way to assess the effect of blood storage on the oxygen transport rate (OTR).
Materials and Methods  In dynamic oximetry, the rate at which oxyhemoglobin becomes deoxyhemoglobin is measured optically, thereby, indirectly measuring the rate at which oxygen leaves the red blood cell (RBC) making it available for transfer to tissues. Extending the physiologic diffusion time in an in vitro apparatus, consisting of a diffusion system and gas exchanger capable of controlling the surface area and the time of exposure for oxygenation and deoxygenation, makes OTR measurement feasible. Eight normal blood donor units, collected in adenine, dextrose, sorbitol, sodium chloride and mannitol , were stored for 8 weeks under standard conditions and serially sampled for OTR.
Results  We report that the OTR at the time of blood bank donation appears to be singular for each donor, that the interdonor differences are maintained over time, and that the individual OTR increased 1·72-fold (95% CI 1·51, 1·95) over 8 weeks, adjusting for sex, age and plasma cholesterol level.
Conclusion  Oxygen transport rate increases during storage; blood units with similar haemoglobin content may have significant differences in OTR. Studies examining blood parameters at the time of donation and blood storage on patient outcomes should consider measuring OTR, as it may contribute to differences in observed efficacy of tissue oxygenation.     

Multicentre verification of haematology laboratory blood collection tubes during a global blood collection tube shortage

Introduction

Verification of blood collection tubes is essential for clinical laboratories. The aim of this study was to assess performance of candidate tubes from four alternative suppliers for routine diagnostic haematology testing during an impending global shortage of blood collection tubes.

Methods

A multicentre verification study was performed in Cape Town, South Africa. Blood from 300 healthy volunteers was collected into K₂EDTA and sodium citrate tubes of BD Vacutainer® comparator tubes and one of four candidate tubes (Vacucare, Vacuette®, V-TUBE™ and Vacutest®). A technical verification was performed, which included tube physical properties and safety. Routine haematology testing was performed for clinical verification.

Results

Vacucare tubes did not have a fill-line indicator, Vacuette® tubes had external blood contamination on the caps post-venesection and Vacutest® tubes had hard rubber stoppers. K₂EDTA tubes of Vacuette®, Vacucare and Vacutest® performed similarly to the comparator. Unacceptable constant bias was seen for PT in Vacucare (95% CI −2.38 to −0.10), Vacutest® (95% CI −1.91 to −0.49) and Vacuette® (95% CI 0.10–1.84) tubes and for aPTT in Vacuette® (95% CI 0.22–2.00) and V-TUBE™ (95% CI −2.88 to −0.44). Unacceptable %bias was seen for aPTT in Vacucare (95% CI 2.78–4.59) and Vacutest® tubes (95% CI 2.53–3.82; desirable ±2.30), and in V-TUBE™ for mean cell volume (95% CI 1.15–1.47, desirable ±0.95%) and mean cell haemoglobin concentration (95% CI −1.65 to −0.93, desirable ±0.43%).

Conclusion

Blood collection tubes introduce variability to routine haematology results. We recommend that laboratories use one brand of tube. Verification of new candidate tubes should be performed to ensure consistency and reliable reporting of results.     

Pilot study of screening for Wilson disease using dried blood spots obtained from children seen at outpatient clinics

Wilson disease (WD) is an autosomal recessive disorder of copper accumulation leading to liver and/or brain damage. In this paper, we describe the results of a pilot study of screening for WD using ceruloplasmin determinations in dried blood samples. Specimens were collected from children aged 1 to 6 years who were seen at local paediatric outpatient clinics in the Miyagi Prefecture. We measured ceruloplasmin (CP) concentrations in 2789 children using an enzyme-linked immunosorbent assay. The mean value was 12.4±3.95 mg/dl blood. Among these children, we identified two (case 1, male, 2 years old; case 2, female, 3 years old) with markedly reduced CP concentrations. Apart from low serum copper concentrations, their biochemical findings were almost normal, as were growth and development. To confirm the diagnosis, we analysed the WD gene and detected A803T/2871delC mutations in case 1 and R778L/G1035V mutations in case 2. We conclude that these children were presymptomatic WD patients. The CP level in dried blood samples from children aged 1 to 6 years appears to be a reliable marker for early detection of WD.