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1.
J Toxicol Sci ; 47(5): 193-199, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35527007

RESUMO

According to ICH S3A Q&A focusing on microsampling, its application should be avoided in main study animals for test drugs that could exacerbate hematological parameters with frequent blood sampling. However, no study has reported the effects of microsampling on toxicity parameters of drugs known to induce hematological toxicity. Therefore, we assessed the toxicological effects of serial microsampling on rats treated with phenacetin as a model drug. In a common 28-day study, 50 µL of microsampling was performed at 6-time points on days 1 to 2 and 7-time points on days 27 to 28 from the jugular vein of Sprague Dawley rats. The study was performed independently by two organizations. The toxicological influence of microsampling was evaluated on body weight, food consumption, hematology, blood clinical chemistry, urine parameters, organ weights, and tissue pathology. Phenacetin treatments induced significant changes of various hematological parameters (including hemoglobin and reticulocytes), some organ weights (including liver and spleen), and some hematology-related pathological parameters in the liver, spleen and bone marrow. Meanwhile, serial microsampling exhibited minimal influence on the assessed parameters, although 20 parameters showed statistical differences mostly at one organization. The current results support the notion that serial 50 µL microsampling from the jugular vein had minimal impacts on overall toxicological profiles even in rats treated with a drug inducing hematological toxicity, but the potential adverse effect on certain parameters could not be fully excluded. Accordingly, this microsampling technique has possibility to be employed even for non-clinical rat toxicity studies using drugs with potentially hematological toxicity.


Assuntos
Coleta de Amostras Sanguíneas , Fenacetina , Animais , Coleta de Amostras Sanguíneas/métodos , Peso Corporal , Veias Jugulares , Fenacetina/toxicidade , Ratos , Ratos Sprague-Dawley , Baço
2.
Clin Lab ; 68(4)2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-35443592

RESUMO

BACKGROUND: We aimed to compare the Sarstedt S-Monovette serum gel tube and the BD (Becton, Dickinson and Company) Serum Separator Tube II (SST II) Advance based on technical specifications and tests results. METHODS: One hundred and twenty volunteers were included in the technical evaluation and 42 of 120 volunteers in the clinical evaluation. Blood was collected into S-Monovette, and SST II. Twelve quality indicators (QI) were determined for technical evaluation. For clinical evaluation, 29 clinical chemistry analytes were analysed simultaneously on a Roche Cobas 6000 c501 (Roche Diagnostics, Mannheim, Germany). Calculations were made using the formula suggested by the EFLM according to the QIs. If the difference between S-Monovette and SST II was < 1%, S-Monovette was considered sufficient for relevant QI. For clinical evaluation, Passing Bablok regression analysis and Bland-Altman plots were used. Desirable bias values for comparison with mean percentage difference (MPD) were obtained from biological variation databases. RESULTS: S-Monovette tubes were found to be suitable for all QIs (difference < 1%). No significant differences were observed in analytes except lactate dehydrogenase (LDH). LDH results (U/L) obtained from the SST II were statistically significantly higher (SST II: 201 ± 42, S-Monovette: 195 ± 35, regression equation was y = 31.4 + 0.8x). The MPD of LDH (2.4%) remained within the desirable bias (3.4%); however, the 95% CI of the MPD of LDH (0.5% - 4.4%) exceeded the desirable bias. CONCLUSIONS: S-Monovette has been deemed appropriate for use in clinical chemistry analysis, as the MPD of LDH and other analytes remained within the bias limits. The LDH was considered sensitive to microhemolysis as a possible reason for the difference in LDH results.


Assuntos
Coleta de Amostras Sanguíneas , Química Clínica , Coleta de Amostras Sanguíneas/métodos , Alemanha , Humanos , Soro
3.
Clin Lab ; 68(4)2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-35443594

RESUMO

BACKGROUND: Vacuum blood collection tubes with separator gels have made biochemical testing considerably more convenient and provided test samples with better quality; however, the separator gel composition is complex, and it is easy to be polluted by some chemical substances that affect the accuracy of the test results if the raw material control is not strict. METHODS: This study reported a case of pseudo elevation of serum calcium. RESULTS: The pseudo elevation of serum calcium was caused by separator gel calcium contamination of the new batch of vacuum tube (s2005030). CONCLUSIONS: We recommend that clinical laboratories verify the performance of each new batch of vacuum blood collection tubes before using them to prevent the occurrence of adverse events, ensure the accuracy of test results, and provide better clinical services.


Assuntos
Coleta de Amostras Sanguíneas , Cálcio , Coleta de Amostras Sanguíneas/métodos , Géis , Humanos
4.
Medicine (Baltimore) ; 101(10): e29025, 2022 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-35451404

RESUMO

ABSTRACT: We aimed to determine the upper and lower cutoff values to simplify the diagnosis of gestational diabetes mellitus (GDM). We investigated the 50-g oral glucose tolerance test (OGTT) results from 1441 pregnancies and identified 423 gravidas who underwent the 100-g OGTT from 2011 to 2019. We collected the results of 50- and 100-g OGTTs. Moreover, we obtained the sum of the 50-g OGTT and 0-hour values, and the sum of those levels and 1-hour values. We determined the upper cutoff at 50-g OGTT, 0-, 1-hour, sum of 50-g OGTT and 0-hour results, and sum of those levels and 1-hour results for the confirmation of GDM. Also, we determined the lower cutoff at these tests for the exclusion of GDM. The upper cutoffs in 50-g OGTT, 0-, 1-hour, the sum of 50-g OGTT and 0-hour were 222, 115, 212, and 315 mg/dL, respectively. The lower cutoffs in 50-g OGTT, 0-, 1-hour, the sum of 50-g OGTT and 0-hour were 131, 65, 151, and 208 mg/dL, respectively. In addition, we discovered that the upper and lower cutoffs in the sum of 50-g OGTT, 0- and 1-hour values were >516 and <373 mg/dL, respectively. We implemented these cutoffs to our study group at 50-g OGTT and 0-, 1-hour of 100-g OGTT. It could omit 2- and 3-hour sampling in 216 gravidas (51.1%). Our approach was able to simplify GDM diagnostic steps in half of our study group.


Assuntos
Diabetes Gestacional , Glicemia , Coleta de Amostras Sanguíneas , Diabetes Gestacional/diagnóstico , Feminino , Teste de Tolerância a Glucose , Humanos , Flebotomia , Gravidez
5.
PLoS One ; 17(4): e0266567, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35385561

RESUMO

Oxidative stress has a major role in disease pathogenesis. However, limited studies have investigated the effect of various sample collection tubes on oxidative biomarkers. The present study aimed to evaluate the effect of different collection tubes on the variation of malondialdehyde (MDA), nitric oxide (NO), total thiol (t-SH), and ferric reducing ability of plasma (FRAP) levels. A total of 35 individuals participated in this study and each collected sample was separated into three different tubes: glass tubes (GTs), plain plastic tubes (PTs), and gel separator tubes (GSTs). The results of PTs and GSTs were compared to those of GTs as the reference tube. The comparison between the means of biomarkers in various tubes indicated that there was no significant difference in MDA results between tubes. In contrast, t-SH and NO content were significantly decreased in GSTs and PTs compared to GTs. However, the Bland-Altman analysis showed an acceptable concordance for the mentioned analytes and the statistically significant differences were not clinically significant for NO, MDA, and t-SH antioxidant parameters. Moreover, the FRAP level was considerably lower in GSTs compared to GTs. Nevertheless, the Bland-Altman analysis showed a high bias percentage for the FRAP assay when using PTs and GSTs. According to the present results, it can be concluded that switching to plastic blood collection tubes or serum separation tubes could influence the FRAP results. However, there was no interference for the interpretation of other antioxidant assays in different types of collection tubes. Hence, it is suggested to use GTs for total antioxidant capacity evaluations, especially the FRAP assay.


Assuntos
Antioxidantes , Coleta de Amostras Sanguíneas , Biomarcadores , Coleta de Amostras Sanguíneas/métodos , Humanos , Estresse Oxidativo , Plásticos
6.
J Proteome Res ; 21(4): 1061-1072, 2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-35271285

RESUMO

Blood derivatives are the biofluids of choice for metabolomic clinical studies since blood can be collected with low invasiveness and is rich in biological information. However, the choice of the blood collection tubes has an undeniable impact on the plasma and serum metabolic content. Here, we compared the metabolomic and lipoprotein profiles of blood samples collected at the same time and place from six healthy volunteers but using different collection tubes (each enrolled volunteer provided multiple blood samples at a distance of a few weeks/months): citrate plasma, EDTA plasma, and serum tubes. All samples were analyzed via nuclear magnetic resonance spectroscopy. Several metabolites showed statistically significant alterations among the three blood matrices, and also metabolites' correlations were shown to be affected. The effects of blood collection tubes on the lipoproteins' profiles are relevant too, but less marked. Overcoming the issue associated with different blood collection tubes is pivotal to scale metabolomics and lipoprotein analysis at the level of epidemiological studies based on samples from multicenter cohorts. We propose a statistical solution, based on regression, that is shown to be efficient in reducing the alterations induced by the different collection tubes for both the metabolomic and lipoprotein profiles.


Assuntos
Plasma , Soro , Coleta de Amostras Sanguíneas/métodos , Ácido Cítrico/metabolismo , Humanos , Metabolômica/métodos , Plasma/química , Soro/química
7.
Environ Monit Assess ; 194(4): 315, 2022 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-35355133

RESUMO

Mercury (Hg) is a global pollutant and a danger to human health. Human biomonitoring of Hg using a dried blood matrix instead of venous blood sampling for exposure assessment is of growing interest. This study aims to develop, validate, and evaluate the application of volumetric absorptive microsampling (VAMS) for Hg biomonitoring in humans. Sampling, drying, and storage conditions were evaluated during method development. Storage in pre-cleaned glass vials after drying for 2 h in a desiccator ensured analyte stability for at least 4 weeks. Sixty-eight paired capillary VAMS and venous blood samples from volunteers in Munich, Germany, were used for method validation. Hg levels in VAMS and venous blood samples were determined by direct mercury analysis. The limits of detection and quantitation for VAMS were 0.18 and 0.61 µg/l, respectively. However, sensitivity could be improved by using two microsamples for analysis instead of one. Hg levels in VAMS samples correlated very well with Hg levels in venous blood samples (R2 = 0.958). Furthermore, VAMS showed a high accuracy (median recovery: 117%) and precision (median relative standard deviation: 8.7%), especially for Hg concentrations above 1.0 µg/l. In fact, accuracy and precision of VAMS improved with increasing Hg concentrations. In conclusion, VAMS in combination with direct mercury analysis is an accurate and viable alternative for human biomonitoring of Hg.


Assuntos
Monitoramento Biológico , Mercúrio , Adulto , Coleta de Amostras Sanguíneas/métodos , Teste em Amostras de Sangue Seco/métodos , Monitoramento Ambiental , Humanos , Espectrometria de Massas em Tandem/métodos
8.
Clin Lab ; 68(3)2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-35254038

RESUMO

BACKGROUND: Over past decades, the instability of parathyroid hormone (PTH) causes great interference for the clinical laboratory. Contradictory results were reported in many reports about storage conditions and suitable blood collection tubes to ensure PTH stability in the pretreatment phase. METHODS: This study recruited 30 participants including 10 healthy persons, 10 hemodialysis, and 10 hyperparathyroidism patients. Five types of blood collection tubes (EDTA-K3 tube, coagulant tube, heparin anticoagulant tube, gel separating tube, and plain tube) were included to determine whether they were suitable as blood-collecting vessels. The time points and conditions for testing samples included less than 2 hours, 4 hours, and 8 hours at room temperature, and, in parallel, 24 hours, 48 hours, and 72 hours in refrigeration. Two different judgement criteria were used to compare the stability of PTH in different blood vessels. RESULTS: Purely statistical analysis showed that 4 types of blood collection tubes could not perform the same storage ability as EDTA-K3 tube at "T0" time point. Plain tube had the largest drop among all types of blood collection tubes. Compared by pairwise t-test, EDTA-K3 tube could maintain intact PTH for 8 hours (p = 0.998) at room temperature and 24 hours (p = 0.053) in refrigeration. When comparing the total change limit (TCL = 18.8%), at room temperature, EDTA-K3 tube (7.0%), heparin tube (12.7%), coagulant tube (16.2%), and plain tube (17.6%) could maintain intact PTH for 8 hours, and GST can preserve PTH for 4 hours (18.2%). In refrigeration, EDTA-K3 tube could maintain PTH for 72 hours (7.5%) and heparin tube could maintain 24 hours (18.4%). The other three blood collection tubes could not preserve PTH in refrigeration (GST = 22.1%, coagulant tube = 20.3%, plain tube = 20.8%). CONCLUSIONS: PTH seems more stable in the EDTA-K3 tube than any other blood collection tubes and is followed next by the heparin anticoagulant tube. Plain tube and GST have faster degradation than other tubes and are not suggested to preserve intact PTH.


Assuntos
Coleta de Amostras Sanguíneas , Hormônio Paratireóideo , Anticoagulantes , Coleta de Amostras Sanguíneas/métodos , Ácido Edético , Testes Hematológicos/métodos , Humanos , Diálise Renal
9.
J Vis Exp ; (180)2022 02 09.
Artigo em Inglês | MEDLINE | ID: mdl-35225291

RESUMO

Rats are widely used in pharmacokinetics (PK) and toxicokinetic (TK) studies that need to collect a certain amount of blood at specific time points to detect drug exposure. The rat blood sampling method determines the quality of the plasma and further affects the precision of the test results. The subclavian vein blood collection method described in this protocol collects blood samples repeatedly in the conscious state of animals to meet the needs of PK and TK tests. The skills of restraint handling and appropriate procedure of needle incision ensure the success rate of blood collection. It is easy to operate while ensuring the quality of plasma and at the same time catering to animal welfare. However, this method requires skilled operation, and an improper one may cause animal weakness, pain, lameness, and even mortality. The current method has been used in the testing facility for a 4-week oral toxicity study in Sprague Dawley (SD) rats with TK. The maximum amount of blood collected within 24 h did not exceed 20% of the animal's total blood. The animals' body weight was more than 200 g for males and females. The data showed that the animals' bodyweight increased steadily every week, and the clinical observation was normal after repetitive sample collection.


Assuntos
Coleta de Amostras Sanguíneas , Veia Subclávia , Bem-Estar do Animal , Animais , Coleta de Amostras Sanguíneas/métodos , Feminino , Masculino , Flebotomia , Ratos , Ratos Sprague-Dawley
10.
Clin Chem Lab Med ; 60(5): 707-713, 2022 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-35167733

RESUMO

OBJECTIVES: Academics are far from a consensus regarding the effects of pneumatic tube system (PTS) delivery on sample integrity and laboratory test results. As for the reasons for conflicting opinions, each PTS is uniquely designed, sample tubes and patient characteristics differ among studies. This study aims to validate the PTS utilized in Ankara City Hospital for routine chemistry, coagulation, and hematology tests by comparing samples delivered via PTS and porter. METHODS: The study comprises 50 healthy volunteers. Blood samples were drawn into three biochemistry, two coagulation, and two hemogram tubes from each participant. Each of the duplicate samples was transferred to the emergency laboratory via Swiss log PTS (aka PTS-immediately) or by a porter. The last of the biochemistry tubes were delivered via the PTS, upon completion of coagulation of the blood (aka PTS-after). The results of the analysis in these groups were compared with multiple statistical analyses. RESULTS: The study did not reveal any correlation between the PTS and serum hemolysis index. There were statistically significant differences in several biochemistry tests. However, none of them reached the clinical significance threshold. Basophil and large unidentified cell (LUC) tests had poor correlations (r=0.47 and r=0.60; respectively) and reached clinical significance threshold (the average percentages of bias, 10.2%, and 15.4%, respectively). The remainder of the hematology and coagulation parameters did not reach clinical significance level either. CONCLUSIONS: The modern PTS validated in this study is safe for sample transportation for routine chemistry, coagulation, and hematology tests frequently requested in healthy individuals except for basophil and LUC.


Assuntos
Coleta de Amostras Sanguíneas , Hematologia , Coagulação Sanguínea , Testes de Coagulação Sanguínea/métodos , Coleta de Amostras Sanguíneas/métodos , Hospitais Urbanos , Humanos
11.
PLoS One ; 17(2): e0264432, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35226681

RESUMO

INTRODUCTION: Sodium fluoride (NaF) tubes are the recommended tubes for glucose measurements, but these are expensive, have limited number of uses, and are not always available in low resource settings. Alternative sample handling techniques are thus needed. We compared glucose stability in samples collected in various tubes exposed to different pre-analytical conditions in Uganda. METHODS: Random (non-fasted) blood samples were drawn from nine healthy participants into NaF, Ethylenediaminetetraacetic acid (EDTA), and plain serum tubes. The samples were kept un-centrifuged or centrifuged with plasma or serum pipetted into aliquots, placed in cool box with ice or at room temperature and were stored in a permanent freezer after 0, 2, 6, 12 and 24 hours post blood draw before glucose analysis. RESULTS: Rapid decline in glucose concentrations was observed when compared to baseline in serum (declined to 64%) and EDTA-plasma (declined to 77%) after 6 hours when samples were un-centrifuged at room temperature whilst NaF-plasma was stable after 24 hours in the same condition. Un-centrifuged EDTA-plasma kept on ice was stable for up to 6 hours but serum was not stable (degraded to 92%) in the same conditions. Early centrifugation prevented glucose decline even at room temperature regardless of the primary tube used with serum, EDTA-plasma and NaF-plasma after 24 hours. CONCLUSION: In low resource settings we recommend use of EDTA tubes placed in cool box with ice and analysed within 6 hours as an alternative to NaF tubes. Alternatively, immediate separation of blood with manual hand centrifuges will allow any tube to be used even in remote settings with no electricity.


Assuntos
Glicemia/metabolismo , Coleta de Amostras Sanguíneas , Ácido Edético/farmacologia , Fluoreto de Sódio/farmacologia , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
12.
Artigo em Inglês | MEDLINE | ID: mdl-35139475

RESUMO

Erythrocyte membrane-incorporated phosphatidylethanol (PEth) forms only in the presence of ethanol and, once formed, provides a persisting marker for historical alcohol consumption. Relationships between PEth concentration, extent of consumption and time from consumption are under investigation. Threshold values of PEth have been proposed as indicators for any, or for harmful alcohol consumption. Here, we describe an assay for erythrocyte PEth 16:0/18:1 that offers the efficiency needed for routine clinical deployment, in the context of a fully validated methodology. However, we observe that conventional procedures for validating assay methodology are insufficient where the analyte of interest, membrane-incorporated PEth 16:0/18:1, has different physicochemical properties to the soluble PEth 16:0/18:1 and PEth 16:0/18:1-d5 that are used for making calibrator, controls and internal standards. Whereas the internal standard did fully correct for differences in matrix effects and recovery when different extraction solvents were applied to calibrators and controls (in soluble form), it failed to correct for a 1.5-fold difference in the relative efficiency of two solvents, in this case, acetonitrile and isopropanol in extracting PEth from erythrocyte membrane in clinical samples. Differences in the efficiency of the extraction of membrane-bound PEth translate to different results from the same specimen. That can mean that threshold values derived by one methodology cannot be safely generalised to another. That hampers the generalisability of individual laboratory's experience with PEth assay results. Harmonising extraction methodology between laboratories becomes very important where membrane-incorporated PEth itself remains unavailable as an assay standard.


Assuntos
Cromatografia Líquida/métodos , Glicerofosfolipídeos/sangue , Espectrometria de Massas em Tandem/métodos , Consumo de Bebidas Alcoólicas , Biomarcadores/sangue , Coleta de Amostras Sanguíneas , Feminino , Humanos , Limite de Detecção , Modelos Lineares , Masculino , Reprodutibilidade dos Testes
13.
J Pharm Biomed Anal ; 211: 114623, 2022 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-35121279

RESUMO

Personalized dosing of kinase inhibitors (KI) might be beneficial in oral anti-cancer therapy to overcome individual pharmacokinetic variability. Volumetric absorptive microsampling (VAMS) has emerged as an attractive alternative compared to conventional invasive sampling methods enabling remote and frequent specimen collection. Therefore, an LC-MS/MS VAMS method was developed and validated to monitor drug exposure of ten KI from 20 µL dried capillary blood. The assay includes the KI cabozantinib, dabrafenib, nilotinib, and osimertinib with a calibration range of 6-1500 ng/mL and afatinib, axitinib, bosutinib, lenvatinib, ruxolitinib and trametinib within a range of 2-500 ng/mL. Using acetonitrile containing isotope labelled internal standards (IS) as solid-liquid extraction solvent, analytes and IS were detected by multiple reaction monitoring (MRM) after electro-spray ionization (ESI) in positive ionization mode after chromatographic separation using a phenyl-hexyl column. The method was validated according to the FDA and EMA guidelines for bioanalytical method validation and in accordance with the guideline of the International Association for Therapeutic Drug Monitoring and Clinical Toxicology for dried blood spot-based methods. The calibration model was linear and reproducible for all KI (R2> 0.994). Furthermore, the validation demonstrated that the VAMS method is accurate, precise, and sensitive. The method fulfilled the acceptance criteria for matrix effects, recovery, carry over, selectivity as well as for the haematocrit effect and all substances proved to be stable in dried condition for at least six weeks at room temperature. In vitro experiments using spiked venous blood were conducted to establish a VAMS-to-plasma conversion factor for each analyte for comparison of VAMS and plasma concentrations. The method was successfully used in a real-life setting demonstrating its applicability in clinical routine. VAMS concentrations of afatinib, cabozantinib, dabrafenib, nilotinib, ruxolitinib and trametinib were assessed in capillary blood samples collected from either trained healthcare professionals or patients at home.


Assuntos
Coleta de Amostras Sanguíneas , Espectrometria de Massas em Tandem , Coleta de Amostras Sanguíneas/métodos , Cromatografia Líquida/métodos , Teste em Amostras de Sangue Seco , Monitoramento de Medicamentos/métodos , Humanos , Espectrometria de Massas em Tandem/métodos
14.
S Afr Med J ; 112(2): 13503, 2022 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-35139991

RESUMO

BACKGROUND: Bloodstream infections are an important cause of mortality in children. Blood cultures (BCs) remain the primary means of identifying organisms and their antibiotic susceptibility profiles. A shortcoming of BCs is that up to 56% of positive cultures will represent contaminants. Poor adherence to standard practices applicable to BC sampling could explain an unacceptable contamination rate. OBJECTIVES: To determine: (i) the BC contamination rate in the departments of paediatrics and child health at two tertiary hospitals in central South Africa; and (ii) BC sampling practices among paediatric clinicians. METHODS: The author determined the prevalence of BC contamination by analysis of laboratory data for the period 1 May - 27 August 2019, and assessed possible factors contributing to BC contamination by surveying paediatric medical staff with a self-administered BC practices questionnaire. RESULTS: Of the 244 BCs reviewed, 25.4% were positive. The most commonly isolated pathogens were coagulase-negative staphylococci (CoNS) (33.3%), Escherichia coli (22.2%), Enterococcus faecium (16.7%) and Acinetobacter baumannii (11.1%). In total, 15.2% of the BCs yielded contaminants and 2.9% had polymicrobial growth. The most common contaminant was CoNS. Approximately 68% of clinicians were not aware of BC sampling guidelines, and even among those who were aware of the guidelines, non-compliance was reported. CONCLUSIONS: The BC contamination rate was higher than internationally accepted rates. Educating clinicians on specific BC sampling guidelines is strongly recommended to decrease the high rate of contamination observed in this study.


Assuntos
Bacteriemia/diagnóstico , Hemocultura/normas , Coleta de Amostras Sanguíneas/normas , Manejo de Espécimes/normas , Bacteriemia/microbiologia , Técnicas Bacteriológicas/normas , Guias como Assunto , Humanos , Pediatria , Prevalência , África do Sul , Inquéritos e Questionários , Centros de Atenção Terciária
15.
BMC Res Notes ; 15(1): 61, 2022 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-35172879

RESUMO

OBJECTIVE: Self-sampling of capillary blood provides easier sample collection, handling, and shipping compared to more invasive blood sampling via venepuncture. Recently, other means of capillary blood collection were introduced to the market, such as Neoteryx sticks and Noviplex cards. We tested the comparability of these two self-sampling methods, alongside dried blood spots (DBS), with plasma acquired from venepunctured blood in N-glycoprofiling of total proteins. We have also tested the intra-day repeatability of the three mentioned self-sampling methods. Capillary blood collection with Neoteryx, Noviplex and DBS was done following the manufacturers' instructions and N-glycoprofiling of released, fluorescently labelled N-glycans was performed with ultra-performance liquid chromatography. RESULTS: Comparability with plasma was assessed by calculating the relative deviance, which was 0.674 for DBS, 0.092 for Neoteryx sticks, and 0.069 for Noviplex cards. In repeatability testing, similar results were obtained, with Noviplex cards and Neoteryx sticks performing substantially better than DBS (CVs = 4.831% and 7.098%, compared to 14.305%, respectively). Our preliminary study on the use of Neoteryx and Noviplex self-sampling devices in glycoanalysis demonstrates their satisfactory performance in both the comparability and repeatability testing, however, they should be further tested in larger collaborations and cohorts.


Assuntos
Teste em Amostras de Sangue Seco , Manejo de Espécimes , Coleta de Amostras Sanguíneas , Cromatografia Líquida , Teste em Amostras de Sangue Seco/métodos , Glicosilação , Manejo de Espécimes/métodos
16.
Scand J Clin Lab Invest ; 82(2): 115-122, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35112638

RESUMO

The analytical stability of laboratory tests relies mostly on internal and external quality control procedures. Summarized patient data has in several studies been shown to be a good supplement for monitoring analytical stability. In our present investigation, we evaluate a datamining method for retrospective evaluation and assessment of analyte stability in whole blood. Results from the laboratory information system were used as the basis for the datamining approach. Blood tests were requested by the general practitioners and drawing of the blood sample was either at the general practitioner's or at the hospital outpatient clinics. We were able to split data into groups based on sample collection place and time to analysis. The datamining approach was compared to experiments where samples were incubated at a single temperature as well as an experiment where the temperatures were changed during incubation. To demonstrate the method, we selected three laboratory tests considered representative: potassium, phosphate, and lactate dehydrogenase. The datamining approach showed results similar to the reference experiment. Furthermore, our results show that the analytes phosphate and potassium were not stable after short storage at a lower temperature.


Assuntos
Coleta de Amostras Sanguíneas , Potássio , Coleta de Amostras Sanguíneas/métodos , Humanos , Fosfatos , Estudos Retrospectivos , Manejo de Espécimes , Temperatura
17.
Clin Chem Lab Med ; 60(5): 689-700, 2022 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-35073617

RESUMO

OBJECTIVES: The use of BD Vacutainer® Barricor™ tubes (BAR) can reduce turnaround time (TAT) and improve separation of plasma from cellular components using a specific mechanical separator. Concentrations of amino acids (AAs) and cytokines, known to be labile during pre-analytical time delays, were compared in heparin (BAR, BD Heparin standard tube [PST]), EDTA and serum gel tubes (SER) to validate previously identified quality indicators (QIs) in BAR. METHODS: Samples of healthy individuals (n=10) were collected in heparin, EDTA and SER tubes and exposed to varying pre- and post-centrifugation delays at room temperature (RT). Cytokines (interleukin [IL]-8, IL-16 and sCD40L) were analyzed by enzyme-linked immunosorbent assay (ELISA) and AAs were characterized by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). RESULTS: All QIs, AAs/AA ratio and cytokines increased during prolonged blood storage in heparin plasma (PST, BAR) and SER tubes. Comparison of 53 h/1 h pre-centrifugation delay resulted in an increase in taurine (Tau) and glutamic acid (Glu) concentrations by more than three times, soluble CD40L increased by 13.6, 9.2 and 4.3 fold in PST, BAR-CTRL and BAR-FAST, and IL-8 increased even more by 112.8 (PST), 266.1 (BAR-CTRL), 268.1 (BAR-FAST) and 70.0 (SER) fold, respectively. Overall, compared to prolonged blood storage, effects of post-centrifugation delays were less pronounced in all tested materials. CONCLUSIONS: BAR tubes are compatible with the use of several established QIs and can therefore be used in clinical biobanking to reduce pre-analytical TAT without compromising QIs and thus pre-analytical sample quality analysis.


Assuntos
Aminoácidos , Citocinas , Bancos de Espécimes Biológicos , Coleta de Amostras Sanguíneas/métodos , Cromatografia Líquida , Humanos , Indicadores de Qualidade em Assistência à Saúde , Espectrometria de Massas em Tandem
18.
J Endocrinol Invest ; 45(5): 981-987, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35098493

RESUMO

BACKGROUND: In patients with suspected acromegaly, evaluation of IGF-I is recommended as first-line test, while the assessment of GH-nadir during oral glucose tolerance test (OGTT) is advised as confirmatory test. The procedure of this test generally involves GH measurement every 30 min (30') from baseline to +120' or +180'. However, the optimal timing of samplings for the distinction between patients with or without active acromegaly is still a matter of debate. METHODS: Sixty-seven healthy subjects and 46 acromegalic patients who achieved documented and persistent long-term cure were enrolled. A greedy algorithm was used to identify the minimal subset of time-points that sufficed to correctly detect GH suppression. RESULTS: The sampling at 90' was the one in which a GH level < 1 µg/L was most frequently achieved (i.e., in 91.3% of cured acromegalic patients and in 91.0% of healthy subjects). Considering the whole cohort, the best combination of 2 time-points was +90' and +150' and achieved 95.6% accuracy; the best combination of 3 time-points was +60', +90' and +150' and achieved 99.1% accuracy. The minimal subset of GH determinations that demonstrated perfect accuracy (100%) needed the inclusion of 4 time-points, namely +60', +90', +120' and +150'. CONCLUSION: A subset of 4 time-points (60' - 90' - 120' - 150') was identified as the most relevant to detect GH suppression at OGTT, with a perfect classification of 100% of subjects. This supports the possibility to restrict the blood samplings to these time-points when assessing disease cure, with possible advantages in terms of saving time and lowering costs.


Assuntos
Acromegalia , Hormônio do Crescimento Humano , Acromegalia/diagnóstico , Coleta de Amostras Sanguíneas , Estudos de Coortes , Teste de Tolerância a Glucose , Humanos , Fator de Crescimento Insulin-Like I
19.
Anal Bioanal Chem ; 414(6): 2155-2162, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34981148

RESUMO

Reducing the volume of blood sampled from neonatal or paediatric patients is important to facilitate research in a group that is under-represented in clinical studies. Not all patients have a cannula available for blood sampling, meaning there are real advantages in obtaining a blood microsample by skin prick. In this study, the results obtained from both capillary microsamples (CMS) and a microfluidic (MF)-CMS by skin prick are compared to conventional plasma sampled from an arterial catheter in a clinical bridging study. Six critically ill patients receiving meropenem were included with the incurred sample reanalysis test meeting the acceptance criteria for both CMS (n = 24 samples) and MF-CMS (n = 20 samples). Bland-Altman plots comparing MF-CMS to conventional arterial blood sampling revealed a difference of - 12.7 ± 22.1% (mean ± standard deviation (SD), and comparing CMS to conventional arterial blood sampling a difference of - 3.4 ± 17.0%. At - 12.7%, the bias between MF-CMS and conventional sampling is greater than the bias found with CMS, although within the limit of acceptability for analytical accuracy (that being ± 15%). Samples collected by skin prick and using CMS produced meropenem concentrations that were comparable to those obtained from conventional arterial catheter sampling. CMS samples were found to be stable when stored in the capillary tube for 24 h at 5 °C or for 4 h at room temperature.


Assuntos
Coleta de Amostras Sanguíneas , Manejo de Espécimes , Coleta de Amostras Sanguíneas/métodos , Criança , Humanos , Recém-Nascido , Meropeném , Plasma
20.
Artigo em Inglês | MEDLINE | ID: mdl-35055694

RESUMO

AIM: The aim of the study was to assess the severity of pain experienced by a newborn during a heel puncture for screening using the Newborn Pain Scale (NIPS), measure the heart rate and compare the effectiveness of non-pharmacological methods of pain control. DESIGN: Randomized clinical trial. No experimental factors. The test was performed during routine screening. Surroundings: Provincial Specialist Hospital in Olsztyn. PATIENTS/PARTICIPANTS: Pain was assessed in 90 full-term newborns. The newborns were rooming in with their mothers in the hospital. INTERVENTIONS: Newborns were divided into three groups. Three different methods of pain relief were used: breastfeeding, 20% glucose administered orally and non-nutritional sucking. MAIN OUTCOME MEASURES: The primary pain outcome was measured using the NIPS and the secondary pain outcome measures (heart rate, oxygen saturation) were measured using a pulse oximeter. RESULTS: During capillary blood sampling from the heel, most newborns, n = 56 (62.2%), experienced no pain or mild discomfort, severe pain occurred in n = 23 (25.6%) and moderate pain occurred in n = 11 (12.2%). No significant statistical differences were found between the degree of pain intensity and the intervention used to minimize the pain p = 0.24. Statistically significant relationships were demonstrated between heart rate variability and the degree of pain intensity (p = 0.01). There were no statistically significant differences between the newborn's pain intensity and the mother's opinion on the effectiveness of breastfeeding in minimizing pain. CONCLUSIONS: This study did not answer the question of which pain management method used during the heel prick was statistically more effective in reducing pain. However, the results indicate that each of the non-pharmacological interventions (breastfeeding, oral glucose dosing and non-nutritive sucking) applied during heel puncture resulted in effective pain management in most of the newborns enrolled in the study. The relationship between heart rate variability and the severity of pain was confirmed. Mothers of newborns in the breastfeeding group were satisfied with the pain relief methods used in the child and the opportunity to console their newborn during painful procedures in a technologically invasive environment.


Assuntos
Coleta de Amostras Sanguíneas , Dor , Coleta de Amostras Sanguíneas/métodos , Criança , Feminino , Calcanhar , Humanos , Recém-Nascido , Dor/diagnóstico , Dor/prevenção & controle , Manejo da Dor/métodos , Medição da Dor
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