Dark brown serum and plasma samples: a case report

This case report describes occurrence of unusual, dark brown coloration of citrate plasma and serum samples in a female 68 years old patient admitted into Emergency department (ED). Patient complained of nausea and vomiting, fever up to 38.9°C, colicky pain in abdomen, diminished urinary output and yellowish skin tone. Her medical history included arterial hypertension, hypothyroidism and facial squamous cell carcinoma. For previous two years, she was treated with tuberculostatic therapy for Mycobacterium avium positive interstitial lung disease. Regular follow-up showed no signs of active disease. Upon admission to ED, complete blood count (CBC) analysis showed low red blood count (RBC) (3.76 x1012/L (reference interval (RI) 3.86 – 5.08 x1012/L)), low haemoglobin (Hb) concentration (111 g/L (RI 119 - 157 g/L)) and low haematocrit (Hct) (0.310 L/L (RI 0.360 – 0.470 L/L)). Biochemistry analytes were high, with foremost lactate dehydrogenase (LD) activity (2900 U/L, RI < 240 U/L). After communication with the clinician, methaemoglobin measured in arterial blood gas sample was reported. Patient was admitted to the Intensive care unit and upon reflex testing of haptoglobin, intravascular haemolysis was confirmed. This case indicates that every case of brown coloration of the serum must be promptly communicated to the clinician. Reflex testing assured timely diagnosis and favourable patient outcome

The knowledge and understanding of preanalytical phase among biomedicine students at the University of Zagreb

Introduction: The educational program for health care personnel is important for reducing preanalytical errors and improving quality of laboratory test results. The aim of our study was to assess the level of knowledge on preanalytical phase in population of biomedicine students through a cross-sectional survey. Materials and methods: A survey was sent to students on penultimate and final year of Faculty of Pharmacy and Biochemistry – study of medical biochemistry (FPB), Faculty of Veterinary Medicine (FVM) and School of Medicine (SM), University of Zagreb, Croatia, using the web tool SurveyMonkey. Survey was composed of demographics and 14 statements regarding the preanalytical phase of laboratory testing. Comparison of frequencies and proportions of correct answers was done with Fisher’s exact test and test of comparison of proportions, respectively. Results: Study included 135 participants, median age 24 (23-40) years. Students from FPB had higher proportion of correct answers (86%) compared to students from other biomedical faculties 62%, P < 0.001. Students from FPB were more conscious of the importance of specimen mixing (P = 0.027), prevalence of preanalytical errors (P = 0.001), impact of hemolysis (P = 0.032) and lipemia interferences (P = 0.010), proper choice of anticoagulants (P = 0.001), transport conditions for ammonia sample (P < 0.001) and order of draw during blood specimen collection (P < 0.001), in comparison with students from SM and FVM. Conclusions: Students from FPB are more conscious of the importance of preanalytical phase of testing in comparison with their colleagues from other biomedical faculties. No difference in knowledge between penultimate and final year of the same faculty was found

Haemolysis and lipemia interfere with resistin and myeloperoxidase BioVendor ELISA assays

Introduction: The aim of our study was to investigate the influence of haemolysis and lipemia on resistin (RES) and myeloperoxidase (MPO) measurement by BioVendor enzyme-linked immunosorbent assays (ELISA). Materials and methods: Blood was taken from healthy volunteers into lithium heparin tubes. Plasma samples were spiked with Lipofundin® emulsion (B. Braun Melsungen AG, Germany) for lipemia interference testing. Haemolysed samples were obtained by drawing aliquots of heparinized blood through a 26 gauge needle. Index of haemolysis (H), lipemia (L) and triglyceride concentration were measured on Abbott Architect c8000. Haemoglobin concentration was measured on Sysmex XN-1000. Concentrations of RES and MPO in all samples were determined with RES and MPO ELISA kits (BioVendor, Czech Republic). All measurements were performed in triplicate. Biases from the native samples were calculated for both analytes and compared with an arbitrary value (e.g. ± 10%). Results: Triglyceride concentration in the investigated samples ranged from 0.57 to 38.23 mmol/L, which corresponds to L index from - 0.01 to 13.77. Haemoglobin concentration in all samples ranged from 0 to 8 g/L which correspond to H index from 0.05 to 8.77. Both MPO and RES showed significant biases at 1 g/L haemoglobin (58.7% and 66.7%, respectively). Also, both MPO and RES showed significant biases at 4.66 mmol/L triglycerides (33.8% and - 12.2%, respectively). Conclusions: Resistin BioVendor assays are affected by haemolysis and lipemia already at low degree of interferent. Haemolysis was found to interfere at 1 g/L haemoglobin for both assays, while lipemia interferes at 4.66 mmol/L of triglycerides

he knowledge and understanding of preanalytical phase among biomedicine students at the University of Zagreb

Introduction: The educational program for health care personnel is important for reducing preanalytical errors and improving quality of laboratory test results. The aim of our study was to assess the level of knowledge on preanalytical phase in population of biomedicine students through a cross-sectional survey. Materials and methods: A survey was sent to students on penultimate and final year of Faculty of Pharmacy and Biochemistry – study of medical biochemistry (FPB), Faculty of Veterinary Medicine (FVM) and School of Medicine (SM), University of Zagreb, Croatia, using the web tool SurveyMonkey. Survey was composed of demographics and 14 statements regarding the preanalytical phase of laboratory testing. Comparison of frequencies and proportions of correct answers was done with Fisher’s exact test and test of comparison of proportions, respectively. Results: Study included 135 participants, median age 24 (23-40) years. Students from FPB had higher proportion of correct answers (86%) compared to students from other biomedical faculties 62%, P < 0.001. Students from FPB were more conscious of the importance of specimen mixing (P = 0.027), prevalence of preanalytical errors (P = 0.001), impact of hemolysis (P = 0.032) and lipemia interferences (P = 0.010), proper choice of anticoagulants (P = 0.001), transport conditions for ammonia sample (P < 0.001) and order of draw during blood specimen collection (P < 0.001), in comparison with students from SM and FVM. Conclusions: Students from FPB are more conscious of the importance of preanalytical phase of testing in comparison with their colleagues from other biomedical faculties. No difference in knowledge between penultimate and final year of the same faculty was found

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Targeted Mass Spectrometry-Based Assays for Relative Quantification of 30 Brain-Related Proteins and Their Clinical Applications

Cerebrospinal fluid (CSF) is a promising clinical sample for identification of novel biomarkers for various neurological disorders. Considering its direct contact with brain tissue, CSF represents a valuable source of brain-related and brain-specific proteins. Multiple sclerosis is an inflammatory, demyelinating neurological disease affecting the central nervous system, and so far there are no diagnostic or prognostic disease specific biomarkers available in the clinic. The primary aim of the present study was to develop a targeted mass spectrometry assay for simultaneous quantification of 30 brain-related proteins in CSF and subsequently to demonstrate assay feasibility in neurological samples derived from multiple sclerosis patients. Our multiplex selected reaction monitoring assay had wide dynamic range (median fold range across peptides = 8.16 × 10³) and high assay reproducibility (median across peptides CV = 4%). Candidate biomarkers were quantified in CSF samples from neurologically healthy individuals (n = 9) and patients diagnosed with clinically isolated syndrome (n = 29) or early multiple sclerosis (n = 15)