Contamination of lithium heparin blood by K2-ethylenediaminetetraacetic acid (EDTA): an experimental evaluation

Introduction:

The contamination of serum or lithium heparin blood with ethylenediaminetetraacetic acid (EDTA) salts may affect accuracy of some critical analytes and jeopardize patient safety. The aim of this study was to evaluate the effect of lithium heparin sample contamination with different amounts of K₂EDTA.

Materials and methods:

Fifteen volunteers were enrolled among the laboratory staff. Two lithium heparin tubes and one K₂EDTA tube were collected from each subject. The lithium-heparin tubes of each subject were pooled and divided in 5 aliquots. The whole blood of K₂EDTA tube was then added in scalar amount to autologous heparinised aliquots, to obtained different degrees of K₂EDTA blood volume contamination (0%; 5%; 13%; 29%; 43%). The following clinical chemistry parameters were then measured in centrifuged aliquots: alanine aminotranspherase (ALT), bilirubin (total), calcium, chloride, creatinine, iron, lactate dehydrogenase (LD), lipase, magnesium, phosphate, potassium, sodium.

Results:

A significant variation starting from 5% K₂EDTA contamination was observed for calcium, chloride, iron, LD, magnesium (all decreased) and potassium (increased). The variation of phosphate and sodium (both increased) was significant after 13% and 29% K₂EDTA contamination, respectively. The values of ALT, bilirubin, creatinine and lipase remained unchanged up to 43% K₂EDTA contamination. When variations were compared with desirable quality specifications, the bias was significant for calcium, chloride, LD, magnesium and potassium (from 5% K₂EDTA contamination), sodium, phosphate and iron (from 29% K₂EDTA contamination).

Conclusions:

The concentration of calcium, magnesium, potassium, chloride and LD appears to be dramatically biased by even modest K₂EDTA contamination (i.e., 5%). The values of iron, phosphate, and sodium are still reliable up to 29% K₂EDTA contamination, whereas ALT, bilirubin, creatinine and lipase appear overall less vulnerable towards K₂EDTA contamination.     

Incorrect order of draw could be mitigate the patient safety: a phlebotomy management case report

Procedures involving phlebotomy are critical for obtaining diagnostic blood specimens and represent a well known and recognized problem, probably among the most important issues in laboratory medicine. The aim of this report is to show spurious hyperkalemia and hypocalcemia due to inadequate phlebotomy procedure. The diagnostic blood specimens were collected from a male outpatient 45 years old, with no clinical complaints. The tubes drawing order were as follows: i) clot activator and gel separator (serum vacuum tube), ii) K₃EDTA, iii) a needleless blood gas dedicated-syringe with 80 I.U. lithium heparin, directly connected to the vacuum tube holder system. The laboratory testing results from serum vacuum tube and dedicated syringe were 4.8 and 8.5 mmol/L for potassium, 2.36 and 1.48 mmol/L for total calcium, respectively. Moreover 0.15 mmol/L of free calcium was observed in dedicated syringe. A new blood collection was performed without K₃EDTA tube. Different results were found for potassium (4.7 and 4.5 mmol/L) and total calcium (2.37 and 2.38 mmol/L) from serum vacuum tube and dedicated syringe, respectively. Also free calcium showed different concentration (1.21 mmol/L) in this new sample when compared with the first blood specimen. Based on this case we do not encourage the laboratory managers training the phlebotomists to insert the dedicated syringes in needle-holder system at the end of all vacuum tubes. To avoid double vein puncture the dedicated syringe for free calcium determination should be inserted immediately after serum tubes before EDTA vacuum tubes.     

关于加强我国高等级生物安全实验室体系规划的思考

近年来,随着禽流感、结核病、埃博拉等疫情的不断发生,全球越来越多的医务工作者和科研人员从事危险病原微生物相关的诊断、检测、研究、开发、生产和教学等活动,而这些活动必须在高等级生物安全实验室内进行。文章通过比较国内外高等级生物安全实验室体系发展现状及特点,分析了我国高等级生物安全实验室体系在整体布局、经费投入、管理和支撑体系建设等方面所存在的问题,提出了完善实验室建设布局、加大资金投入、加强管理和支撑体系建设、强化信息和资源共享等建议以推动我国高等级生物安全实验室体系建设,为我国应对新发和突发传染病疫情提供支持。     

实施国家高级别生物安全实验室规划提高生物安全平台保障能力

新冠肺炎疫情不仅给全球卫生安全体系带来了重大考验,也给全球治理结构和格局带来新的冲击;同时,还暴露出全球及我国在公共卫生应急体系和生物安全体系建设等方面的一些不足。生物安全是国家安全的重要组成部分,已成为国家安全的新疆域。其中,高级别生物实验室是开展生物安全科学研究和体现国家安全能力的核心平台。因此,我国需要进一步实施高级别生物安全实验室体系建设规划,形成布局合理、定位明确、功能齐全、分工协作、网络化运行的高级别生物安全实验室国家体系,建设临床、生产、科研活动有机融合的国家生物安全协作中心。     

高校检测实验室质量控制关键要素与管理体系建设方法——北京航空航天大学高性能碳纤维检测评价中心实例分析

高校检测实验室在仪器设备、人员素质、开放性等方面具有独特优势,为保证检测数据准确、客观,建立完善的实验室质量管理体系,实现检测质量有效控制至关重要。在总结分析我国高校检测实验室特点和管理体系中存在问题的基础上,针对实验室认可制度,结合北京航空航天大学碳纤维检测评价中心的工作,提出高校检测实验室质量管理体系的建设方法,阐述检测质量控制的关键要素,总结相关经验。     

Clinical biochemistry laboratory rejection rates due to various types of preanalytical errors

Introduction:

Preanalytical errors, along the process from the beginning of test requests to the admissions of the specimens to the laboratory, cause the rejection of samples. The aim of this study was to better explain the reasons of rejected samples, regarding to their rates in certain test groups in our laboratory.

Materials and methods:

This preliminary study was designed on the rejected samples in one-year period, based on the rates and types of inappropriateness. Test requests and blood samples of clinical chemistry, immunoassay, hematology, glycated hemoglobin, coagulation and erythrocyte sedimentation rate test units were evaluated. Types of inappropriateness were evaluated as follows: improperly labelled samples, hemolysed, clotted specimen, insufficient volume of specimen and total request errors.

Results:

A total of 5,183,582 test requests from 1,035,743 blood collection tubes were considered. The total rejection rate was 0.65 %. The rejection rate of coagulation group was significantly higher (2.28%) than the other test groups (P < 0.001) including insufficient volume of specimen error rate as 1.38%. Rejection rates of hemolysis, clotted specimen and insufficient volume of sample error were found to be 8%, 24% and 34%, respectively. Total request errors, particularly, for unintelligible requests were 32% of the total for inpatients.

Conclusions:

The errors were especially attributable to unintelligible requests of inappropriate test requests, improperly labelled samples for inpatients and blood drawing errors especially due to insufficient volume of specimens in a coagulation test group. Further studies should be performed after corrective and preventive actions to detect a possible decrease in rejecting samples.     

The effect of storage time and freeze-thaw cycles on the stability of serum samples

Introduction:

Optimal storage of serum specimens in central laboratories for a long period for multicenter reference interval studies, or epidemiologic studies remains to be determined. We aimed to examine the analytical stability of chemistry analytes following numerous freeze-thaw and long term storage.

Materials and methods:

Serum samples were obtained from 15 patients. Following baseline measurement, sera of each subject were aliquoted and stored at −20 °C for two experiments. A group of sera were kept frozen for up to 1, 2 and 3 months and then analyzed for stability. The other experiment consisted of one to ten times of freeze and thaw cycles. Total of 17 chemistry analytes were assayed at each time point. The results were compared with those obtained from the initial analysis of fresh samples. Median or mean changes from baseline (T₀) concentrations were evaluated both statistically and clinically according to the desirable bias.

Results:

Of the analytes studied, aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), gamma-glutamyl transferase (GGT), direct bilirubin, glucose, creatinine, cholesterol, triglycerides, high density lipoprotein (HDL) were stable in all conditions. Blood urea nitrogen (BUN), uric acid, total protein, albumin, total bilirubin, calcium, lactate dehydrogenase (LD) were changed significantly (P < 0.005).

Conclusions:

As a result, common clinical chemistry analytes, with considering the variability of unstable analytes, showed adequote stability after 3 months of storage in sera at −20 °C, or up to ten times of freeze-thaw cycle. All the same, such analysis can only be performed for exceptional cases, and this should be taken into account while planning studies.     

Exhaled breath condensate – from an analytical point of view

Over the past three decades, the goal of many researchers is analysis of exhaled breath condensate (EBC) as noninvasively obtained sample. A total quality in laboratory diagnostic processes in EBC analysis was investigated: pre-analytical (formation, collection, storage of EBC), analytical (sensitivity of applied methods, standardization) and post-analytical (interpretation of results) phases. EBC analysis is still used as a research tool. Limitations referred to pre-analytical, analytical, and post-analytical phases of EBC analysis are numerous, e.g. low concentrations of EBC constituents, single-analyte methods lack in sensitivity, and multi-analyte has not been fully explored, and reference values are not established. When all, pre-analytical, analytical and post-analytical requirements are met, EBC biomarkers as well as biomarker patterns can be selected and EBC analysis can hopefully be used in clinical practice, in both, the diagnosis and in the longitudinal follow-up of patients, resulting in better outcome of disease.     

Effect of sample type,centrifugation and storage conditions on vitamin D concentration

Introduction:

Studies about vitamin D [25(OH)D] stability in plasma are limited and preanalytical variables such as tube type may affect results. We aimed to evaluate effect of storage conditions, sample type and some preanalytical variables on vitamin D concentration.

Materials and methods:

Blood samples from 15 healthy subjects were centrifuged at different temperatures and stored under different conditions. Serum and plasma 25(OH)D difference, effect of centrifugation temperature and common storage conditions were investigated.

Results:

There was no difference between serum and plasma vitamin D concentration. Centrifugation temperature had no impact on vitamin D concentration. 25(OH)D is stable under common storage conditions: 4 hours at room temperature, 24 hours at 2–8 °C, 7 days at −20 °C, 3 months at −80 °C.

Conclusion:

Vitamin D does not require any special storage conditions and refrigeration. Both serum and plasma can be used for measurement.     

Preanalytical quality improvement – in quality we trust

Background

The preanalytical phase represents the major source of variability in laboratory diagnostics. Our aim was to assess to what extent underfilling of primary blood tubes may impact upon routine coagulation testing.

Materials and methods:

Blood was drawn by syringe from 21 healthy volunteers and 6 patients on warfarin therapy, and immediately transferred into 3.6 mL vacuum tubes containing 3.2% sodium citrate (Terumo Europe N.V., Leuven, Belgium). All tubes were filled using standardized volumes of whole blood to produce scalar amounts of filling: 3.6 mL (i.e., 100%), 3.2 mL (89%), 2.8 mL (78%) and 2.4 mL (67%). Samples were mixed and centrifuged at 1300 × g for 10 min. The plasma was tested for prothrombin time (PT), activated partial thromboplastin time (APTT) and fibrinogen (FBG) on ACL TOP (Instrumentation Laboratory - IL, Milan, Italy), using IL reagents. A polynomial plot was derived for each parameter from interpolation of clotting values obtained with different percentages of filling, and these plots were compared with quality specifications (± 2.0 for PT, ± 2.3 for APTT and ± 4.8 for FBG) to calculate the minimal filling volume required to produce clinically acceptable results.

Results:

The equations were (PF, Percentage of filling): PT (sec) = 3.375 × PF^2–6.255 × PF + 17.806 (r = 0.980); APTT (sec) = 8.925 × PF^2–23.578 × PF + 46.356 (r = 0.979); and FBG (mg/dL) = −311.5 × PF^2 + 422.1 × PF + 147.07 (r = 0.994). According to these equations, the minimum allowed thresholds of blood tubes filling were > 61% for PT, > 87% for APTT and > 71% for FBG.

Conclusions:

Our results confirm that routine coagulation testing performed on underfilled tubes may generate biased and clinically misleading test results. This is particularly critical for APTT, wherein tubes filled at less than ∼90% generate unreliable data. The FBG and the PT seem more resistant to underfilling, clinical significant biases being observed only where blood tubes were filled at less than ∼60 and ∼70%, respectively.