Assessment of antinuclear antibodies (ANA): National recommendations on behalf of the Croatian society of medical biochemistry and laboratory medicine

Antinuclear antibodies (ANA) represent a family of autoantibodies targeting ubiquitous cellular constituents and are a hallmark of systemic inflammatory autoimmune rheumatic diseases named connective tissue diseases (CTD). The gold standard method for ANA determination is indirect immunofluorescence (IIF) on the human laryngeal epidermoid carcinoma cell line type 2 substrate (HEp-2), but with increasing demand for ANA testing, novel methods eased for automation emerged, which allows testing by staff less experienced in this specific field of laboratory diagnostic. In 2016 The working group (WG) for laboratory diagnostics of autoimmune diseases as part of the Committee for the Scientific Professional Development of the Croatian Society of Medical Biochemistry and Laboratory Medicine (CSMBLM) published the data of a survey regarding general practice in laboratory diagnostics of autoimmune diseases in Croatia. Results indicated high diversity in the performance of autoantibody testing as well as reporting of the results and indicated the need of creating recommendations for the assessment of ANA that would help harmonize diagnostics of systemic autoimmune rheumatic diseases in Croatia. This document encompasses twenty-seven recommendations for ANA testing created concerning indications for ANA testing, preanalytical, analytical, and postanalytical issues, including rational algorithm and quality control assurance. These recommendations are based on the relevant international recommendations and guidelines for the assessment of ANA testing and relevant literature search and should help to harmonize the approach in ANA testing and clarify differences in interpretation of the results obtained using different methods of determination

Red blood cell distribution width as a prognostic marker of mortality in patients on chronic dialysis: a single center, prospective longitudinal study

Aim To determine if red cell distribution width (RDW) is associated with all-cause mortality in patients on chronic dialysis and to evaluate its prognostic value among validated prognostic biomarkers. Methods This is a single center, prospective longitudinal study. At the time of inclusion in January 2011, all patients were physically examined and a routine blood analysis was performed. A sera sample was preserved for determination of NT-pro-brain natriuretic peptide (NT-pro-BNP) and eosinophil cationic protein. Carotid intima media thickness (IMT) was also measured. Following one year, all-cause mortality was evaluated. Results Of 100 patients, 25 patients died during the follow- up period of one-year. Patients who died had significantly higher median [range] RDW levels (16.7% [14.3-19.5] vs 15.5% [13.2-19.7], P < 0.001. They had significantly higher Eastern Cooperative Oncology Group (ECOG) performance status (4 [2-4] vs 2 [1-4], P < 0.001), increased intima-media thickness (IMT) (0.71 [0.47-1.25] vs 0.63 [0.31-1.55], P = 0.011), increased NT-pro-BNP levels (8300 [1108-35000] vs 4837 [413-35000], P = 0.043), and increased C-reactive protein (CRP) levels (11.6 [1.3-154.2] vs 4.9 [0.4-92.9], P < 0.001). For each 1% point increase in RDW level as a continuous variable, one-year all cause mortality risk was increased by 54% in univariate Cox proportional hazard analysis. In the final model, when RDW was entered as a categorical variable, mortality risk was significantly increased (hazard ratio, 5.15, 95% confidence interval, 2.33 to 11.36) and patients with RDW levels above 15.75% had significantly shorter survival time (Log rank P < 0.001) than others. Conclusions RDW could be an additive predictor for allcause mortality in patients on chronic dialysis. Furthermore, RDW combined with sound clinical judgment improves identification of patients who are at increased risk compared to RDW alone

Comparability of Pathromtin SL, Dade Actin FS i STA Cephascreen reagens for activated partial thromboplastin time measurement

Cilj: Ispitati korelaciju vrijednosti APTV dobivenih korištenjem triju reagensa: Pathromtin SL i Dade Actin FS na analizatoru Berichrom Coagulation System (BCS), odnosno STA Cephascreen na analizatoru STA Compact. Materijali i metode: APTV je određen u 114 uzoraka svježe plazme ispitanika, od kojih je 46 liječeno niskomolekularnim heparinom, a ostalih 68 nije bilo na antikoagulantnoj terapiji. Vrijednosti APTV određene su koagu-lometrijskom metodom usporedno pomoću sva tri reagensa neposredno nakon donošenja uzoraka u koagulacijski laboratorij. Vrijednosti APTV određene su i u komercijalnim kontrolnim pripravcima Lypocheck Coagulation Control proizvođača Bio-Rad (Bio-Rad Laboratories, SAD), koji obuhvaćaju tri različita raspona vrijednosti. Rezultati: Vrijednosti komercijalnih kontrolnih uzoraka Bio-Rad kretale su se unutar raspona što ga navodi proizvođač za sva tri ispitivana reagensa. Korelacijom reagensa Pathromtin SL i Dade Actin FS, Pathromtin SL i STA Cephascreen, te Dade Actin FS i STA Cephascreen dobivene su izvrsne povezanosti (R = 0,9485,0,9353 i 0,9072). Ispitana je razlika između dobivenih koeficijenata korelacije i nađena je statistički značajna razlika između koeficijenata korelacije rezultata dobivenih reagensima Pathromtin SL i Dade Actin FS. Passing-Bab-lok regresijom vrijednosti nagiba i odsječka na osi y obuhvaćale su 1 odnosno 0 samo u kombinaciji reagensa Dade Actin FS i STA Cephascreen. Zaključak: Ispitivanje je pokazalo da su najbolje usporedive vrijednosti dobivene reagensima Dade Actin FS i STA Cephascreen. Unatoč izvrsnoj povezanosti Passing-Bablok regresija pokazala je da su reagensi Pathromtin SL i Dade Actin FS te Pathromtin SL i STA Cephascreen slabije usporedivi. Kako dobiveni rezultati potvrđuju opažanja iz svakodnevnog rada, bitno je da se bolesnici izloženi heparinskom liječenju ne prate istodobno različitim reagensima zbog različite osjetljivosti reagensa prema terapiji niskomolekularnim heparinom.Aim: Aim of the study was to investigate correlation of activated partial thromboplastin time (APTT) measured with three different reagents: Pathromtin SLand Dade Actin FS reagents on Berichrom Coagulation System analyzer, and by STA Cephascreen reagent on STA Compact analyzer. Material and methods: APTT was determined in parallel by Pathromtin SL and Dade Actin FS reagents, and by STA Cephascreen reagent in 114 fresh plasma samples from subjects administered low-molecular-weight heparin and in subjects known to receive no anticoagulant therapy. APTT values were determined by coagulometry method immediately upon the material receipt at coagulation laboratory. APTT values were also determined in Bio-Rad Lypocheck Coagulation Control samples (Bio-Rad Laboratories, USA) covering three different value ranges. Results: The values recorded in Bio-Rad controls were within the range declared by the manufacturer for all three reagents used. Correlation of APTT values obtained by use of Pathromtin SL vs. Dade Actin FS, Pathromtin SL vs. Cephascreen and Dade Actin FS vs. STA Cephascreen yielded strong correlations (R = 0.9485,0.9353 and 0.9072, respectively). A statistically significant difference was recorded between the results obtained by Pathromtin SL and Dade Actin FS reagents. Passing-Bablok regression slope and y-axis intercept included 1 and 0 only for Dade Actin FS vs. STA Cephascreen. Conclusion: Study results showed the best compliance of values obtained by Dade Actin FS and STA Cephascreen reagents. In spite of strong correlations, Passing-Bablok regression indicated lower comparability between Pathromtin SL and Dade Actin as well as between Pathromtin SL and STA Cephascreen reagents. As the study results confirmed the observations from daily routine, it is of utmost importance that individual patients receiving heparin therapy be not monitored by use of different reagents due to the variable reagent sensitivity to low molecular weight heparin

Oxidative Stress Markers in Patients with Post-Traumatic Stress Disorder

Recent study data support the role of oxidative stress in diverse psychiatric disorders. Oxidative stress results from an oxidant/antioxidant imbalance, an excess of oxidants and/or a depletion of antioxidants. There are numerous studies that indicate that free radicals (FRs) damage neurons, and then play an important role in the pathophysiology of schizophrenia and depression. Active oxygen can cause considerable damage and disrupt the important physiological functions of proteins, lipids, enzymes and DNA. The aim of our study was to investigate the possible differences in the concentration of tromboxane B2, 8-OHdG and protein carbonyls, as significant markers of oxidative damage, and urate, albumin and total protein concentrations as antioxidative molecules in PTSD patients in comparison to the healthy control group. The study included 74 male participants who were active soldiers in the Croatian armed forces from 1991 to 1995. 46 subjects with chronic and current PTSD were recruited from the Department of Psychiatry of Dubrava University Hospital during 2010, 28 healthy subjects were recruited in the same period during the regular medical examination at the Dubrava University Hospital. Study results have shown that there is no statistically significant difference in urinary concentrations of 8-OHdG, serum thromboxane B2, and serum urates between two studied groups. Statistically significant difference of the protein carbonyl concentrations was examined. Concentrations were significantly lower in the PTSD group than in the control group. The clinical significance of these results was examined using ROC analysis. The obtained ROC curves did not separate the groups in a satisfactory manner

Oxidative Stress Markers in Patients with Post-Traumatic Stress Disorder

Recent study data support the role of oxidative stress in diverse psychiatric disorders. Oxidative stress results from an oxidant/antioxidant imbalance, an excess of oxidants and/or a depletion of antioxidants. There are numerous studies that indicate that free radicals (FRs) damage neurons, and then play an important role in the pathophysiology of schizophrenia and depression. Active oxygen can cause considerable damage and disrupt the important physiological functions of proteins, lipids, enzymes and DNA. The aim of our study was to investigate the possible differences in the concentration of tromboxane B2, 8-OHdG and protein carbonyls, as significant markers of oxidative damage, and urate, albumin and total protein concentrations as antioxidative molecules in PTSD patients in comparison to the healthy control group. The study included 74 male participants who were active soldiers in the Croatian armed forces from 1991 to 1995. 46 subjects with chronic and current PTSD were recruited from the Department of Psychiatry of Dubrava University Hospital during 2010, 28 healthy subjects were recruited in the same period during the regular medical examination at the Dubrava University Hospital. Study results have shown that there is no statistically significant difference in urinary concentrations of 8-OHdG, serum thromboxane B2, and serum urates between two studied groups. Statistically significant difference of the protein carbonyl concentrations was examined. Concentrations were significantly lower in the PTSD group than in the control group. The clinical significance of these results was examined using ROC analysis. The obtained ROC curves did not separate the groups in a satisfactory manner

Laboratory professionals’ attitudes towards ISO 15189:2012 accreditation: an anonymous survey of three Croatian accredited medical laboratories

Effective implementation and continual compliance with ISO 15189:2012 require ongoing commitment and active involvement of laboratory staff. Our aim was to assess attitudes regarding accreditation implementation by conducting a survey in three Croatian accredited medical laboratories. An anonymous survey consisting of 34 questions was distributed either electronically or in a paper form a week prior to scheduled annual audits. Distributions of answers regarding age, work experience, laboratory workplace, and education level and according to the respective laboratory were compared. The overall response rate was 76% (225/297). Preference towards working in an accredited laboratory and a positive attitude were revealed by 70% and 56% participants, respectively, with better process documentation as the main advantage. Only 14% of responders considered themselves completely familiar with ISO 15189:2012. Total of 68% of responders felt that accreditation increases the usual workload, with excessive paperwork as the main contributor. Half of the responders declared partial agreement that accreditation requirements and expectations were clearly explained and claimed that their suggestions were taken into account only occasionally, which was especially emphasized by technical staff. The vast majority (89%) completely follow the prescribed protocols. Only 27% consider turnaround time monitoring useful. Competence assessment is considered efficient by 41% of responders. The majority (73%) prefer an online audit in times of COVID-19. Despite an overall positive attitude towards accreditation, further efforts are needed in providing better education about ISO 15189:2012 for technical staff and modifying formats of competence assessment, in order to achieve better adherence to ISO 15189:2012 requirements

Comparison of two immunoassays for CA19-9, CEA and AFP tumor markers

Uvod: Monoklonska antitijela koriste se za otkrivanje antigena u serumu koji su povezani sa specifičnim zloćudnim oboljenjima. Ti su tumorski biljezi najkorisniji za praćenje odgovora na terapiju i otkrivanje ranog recidiva, međutim, rezultati dobiveni različitima analizama razlikuju se te promjena metode tijekom praćenja može biti uzrokom problema. Cilj ove studije bio je provesti analitičku evaluaciju usporedivosti analiza za tumorske biljega CA19-9, CEA i AFP na dva različita automatizirana kemijska analizatora. Materijali i metode: Koncentracije CA19-9, CEA i AFP su određene na ana-lizatorima Vitros ECi (Ortho Clinical Diagnostics, Johnson & Johnson, Buckinghamshire, V. Britanija) i Cobase 411 (Hitachi High Technologies Corporation, Tokyo, Japan). Korelacija među metodama ispitana je na 38 uzoraka seruma za CA19-9 i AFP te 39 uzoraka seruma za CEA. Rezultati: Vrijednosti komercijalnih kontrolnih uzoraka bile su unutar raspo-na navedenih od proizvođača za sve tumorske biljege uključene u istraživanje na oba analizatora. Najveće odstupanje od deklariranih kontrolnih vrijednosti nađene su za CA19-9 na analizatoru Vitros ECi te za AFP na analizatoru Cobas e411. Visoka je korelacija utvrđena među metodama za sva tri ispitana tumorska biljega (r = 0,978 za CA19-9; r = 0,995 za CEA, te r = 0,999 za AFP). Nagib i odsječ ak na osi Y iznosili su 1, odnosno 0, samo za usporedbenu analizu AFP. Zaključak: Rezultati istraživanja pokazali su najbolje podudaranje vrijednosti za AFP dobivene na dva ispitana analizatora za imunoanalize. Bez obzira na snažne korelacije, regresija po Passing-Babloku ukazala je na nižu usporedivost dviju imunoanaliza za CEA i CA19-9. S obzirom da su rezultati studije potvrdili zapažanja iz svakodnevnog rutinskog rada, za svakoga je pacijenta od najveće važnosti da ga se prati primjenom iste imunoanalize i reagensa na istom analizatoru.Introduction: Monoclonal antibodies are used to detect serum antigens associated with specific malignancies. These tumor markers are most useful for monitoring response to therapy and detecting early relapse, but results obtained by different assays vary, and a change of method during follow-up may cause problems. The aim of our study was to perform the analytical evaluation of the inter-assay comparability for tumor markers CA19-9, CEA and AFP on two different automated chemistry analyzers. Materials and methods: CA19-9, CEA and AFP concentrations, using Vitros ECi (Ortho Clinical Diagnostics, Johnson and Johnson, Buckinghamshire, UK), and Cobas e 411 (Hitachi High Technologies Corporation, Tokyo, Japan) immu-noassay analyzers, were determined. Between-method correlation was studied in 38 serum samples for CA19-9 and AFP and 39 serum samples for CEA. Results: The values of commercial controls were within the range declared by the manufacturer for all tumor markers included in the study on both analyzers. The highest deviation from the declared control values was found for CA19-9 on Vitros ECi and for AFP on Cobas e411 analyzer. High correlation was found between methods for all of three tumor markers studied (R = 0.978 for CA19-9; R = 0.995 for CEA and R = 0.999 for AFP). Passing-Bablok regression slope and y-axis intercept included 1 and 0 only for AFP comparative assays. Conclusion: Study results showed the best compliance of values for AFP obtained on two studied immunoassay analyzers. Regardless of high correlations, Passing-Bablok regression indicated lower comparability between two immunoassays for CEA and CA19-9. As the study results confirmed the observations from daily routine, it is of utmost importance for individual patients to be monitored using the same immunoassay and reagents on the same analyzer

Comparison of two immunoassays for CA19-9, CEA and AFP tumor markers

Uvod: Monoklonska antitijela koriste se za otkrivanje antigena u serumu koji su povezani sa specifičnim zloćudnim oboljenjima. Ti su tumorski biljezi najkorisniji za praćenje odgovora na terapiju i otkrivanje ranog recidiva, međutim, rezultati dobiveni različitima analizama razlikuju se te promjena metode tijekom praćenja može biti uzrokom problema. Cilj ove studije bio je provesti analitičku evaluaciju usporedivosti analiza za tumorske biljega CA19-9, CEA i AFP na dva različita automatizirana kemijska analizatora. Materijali i metode: Koncentracije CA19-9, CEA i AFP su određene na ana-lizatorima Vitros ECi (Ortho Clinical Diagnostics, Johnson & Johnson, Buckinghamshire, V. Britanija) i Cobase 411 (Hitachi High Technologies Corporation, Tokyo, Japan). Korelacija među metodama ispitana je na 38 uzoraka seruma za CA19-9 i AFP te 39 uzoraka seruma za CEA. Rezultati: Vrijednosti komercijalnih kontrolnih uzoraka bile su unutar raspo-na navedenih od proizvođača za sve tumorske biljege uključene u istraživanje na oba analizatora. Najveće odstupanje od deklariranih kontrolnih vrijednosti nađene su za CA19-9 na analizatoru Vitros ECi te za AFP na analizatoru Cobas e411. Visoka je korelacija utvrđena među metodama za sva tri ispitana tumorska biljega (r = 0,978 za CA19-9; r = 0,995 za CEA, te r = 0,999 za AFP). Nagib i odsječ ak na osi Y iznosili su 1, odnosno 0, samo za usporedbenu analizu AFP. Zaključak: Rezultati istraživanja pokazali su najbolje podudaranje vrijednosti za AFP dobivene na dva ispitana analizatora za imunoanalize. Bez obzira na snažne korelacije, regresija po Passing-Babloku ukazala je na nižu usporedivost dviju imunoanaliza za CEA i CA19-9. S obzirom da su rezultati studije potvrdili zapažanja iz svakodnevnog rutinskog rada, za svakoga je pacijenta od najveće važnosti da ga se prati primjenom iste imunoanalize i reagensa na istom analizatoru.Introduction: Monoclonal antibodies are used to detect serum antigens associated with specific malignancies. These tumor markers are most useful for monitoring response to therapy and detecting early relapse, but results obtained by different assays vary, and a change of method during follow-up may cause problems. The aim of our study was to perform the analytical evaluation of the inter-assay comparability for tumor markers CA19-9, CEA and AFP on two different automated chemistry analyzers. Materials and methods: CA19-9, CEA and AFP concentrations, using Vitros ECi (Ortho Clinical Diagnostics, Johnson and Johnson, Buckinghamshire, UK), and Cobas e 411 (Hitachi High Technologies Corporation, Tokyo, Japan) immu-noassay analyzers, were determined. Between-method correlation was studied in 38 serum samples for CA19-9 and AFP and 39 serum samples for CEA. Results: The values of commercial controls were within the range declared by the manufacturer for all tumor markers included in the study on both analyzers. The highest deviation from the declared control values was found for CA19-9 on Vitros ECi and for AFP on Cobas e411 analyzer. High correlation was found between methods for all of three tumor markers studied (R = 0.978 for CA19-9; R = 0.995 for CEA and R = 0.999 for AFP). Passing-Bablok regression slope and y-axis intercept included 1 and 0 only for AFP comparative assays. Conclusion: Study results showed the best compliance of values for AFP obtained on two studied immunoassay analyzers. Regardless of high correlations, Passing-Bablok regression indicated lower comparability between two immunoassays for CEA and CA19-9. As the study results confirmed the observations from daily routine, it is of utmost importance for individual patients to be monitored using the same immunoassay and reagents on the same analyzer

Red cell distribution width is a potent prognostic parameter for in-hospital and post-discharge mortality in hospitalized coronavirus disease 2019 patients: a registry-based cohort study on 3941 patients

Aim To investigate clinical and prognostic associations of red cell distribution width (RDW) in hospitalized coronavi - rus disease 2019 (COVID-19) patients. Methods We retrospectively analyzed the records of 3941 consecutive COVID-19 patients admitted to a tertiary-level institution from March 2020 to March 2021 who had avail - able RDW on admission. Results The median age was 74 years. The median Charl - son comorbidity index (CCI) was 4. The majority of pa - tients (84.1%) on admission presented with severe or criti - cal COVID-19. Patients with higher RDW were significantly more likely to be older and female, to present earlier dur - ing infection, and to have higher comorbidity burden, worse functional status, and critical presentation of COVID-19 on admission. RDW was not significantly associated with C-re - active protein, occurrence of pneumonia, or need for oxy - gen supplementation on admission. During hospital stay, patients with higher RDW were significantly more likely to require high-flow oxygen therapy, mechanical ventilation, intensive care unit, and to experience prolonged immobi - lization, venous thromboembolism, bleeding, and bacte - rial sepsis. Thirty-day and post-hospital discharge mortality gradually increased with each rising RDW percent-point. In a series of multivariate Cox-regression models, RDW demon - strated robust prognostic properties at >14% cut-off level. This cut-off was associated with inferior 30-day and postdischarge survival independently of COVID-19 severity, age, and CCI; and with 30-day survival independently of COVID severity and established prognostic scores (CURB-65, 4Cmortality, COVID-gram and VACO-index). Conclusion RDW has a complex relationship with COVID19-associated inflammatory state and is affected by prior comorbidities. RDW can improve the prognostication in hospitalized COVID-19 patients