Pre-analytical errors in coagulation testing: a case series.

OBJECTIVES: Prevention of pre-analytical issues in coagulation testing is of paramount importance for good laboratory performance. In addition to common issues like hemolysed, icteric, or lipemic samples, some specific pre-analytical errors of coagulation testing include clotted specimens, improper blood-to-anticoagulant ratio, contamination with other anticoagulants, etc. Prothrombin time (PT) and activated partial thromboplastin time (aPTT) are very commonly affected tests due to pre-analytical variables. The impact these parameters possess on surgical decision-making and various life-saving interventions are substantial therefore we cannot afford laxity and casual mistakes in carrying out these critical investigations at all. CASE PRESENTATION: In this case series, a total of 4 cases of unexpectedly deranged coagulation profiles have been described which were reported incorrectly due to the overall casual approach towards these critical investigations. We have also mentioned how the treating clinician and lab physician retrospectively accessed relevant information in the nick of time to bring back reassurance. CONCLUSIONS: Like every other critical investigation, analytical errors can occur in coagulation parameters due to various avoidable pre-analytical variables. The release of spurious results for coagulation parameters sets alarm bells ringing causing much agony to the treating doctor and patient. Only a disciplined and careful approach taken by hospital and lab staff towards each sample regardless of its criticality can negate these stressful errors to a large extent.

The Role of Indirect Immunofluorescence (IIF) and Line Immunoassay (LIA) in the Diagnosis of Autoimmune Diseases and Their Clinical Correlation: An Observational Study From a Tertiary Care Center in Bihar.

Background and aim The presence of distinct sets of autoantigens and autoantibodies bestow these autoimmune diseases (ADs) with specific immune profiles or fingerprints, which has cleared the diagnostic dilemma associated with these ADs. This study was planned to collate and compare the reporting of indirect immunofluorescence (IIF) with line immunoassay (LIA) and their clinical correlations. This study was conducted to investigate the association between the reporting of anti-nuclear antibody (ANA) screening by IIF and ANA profile reporting by LIA. Additionally, it aimed to explore the association of ANA pattern detection by IIF with the detection of autoantibodies against nuclear antigens by LIA and the association of autoantibody detection by LIA with clinical diagnosis. Methodology A total of 98 samples from patients suspected of having ADs were subjected to both IIF and LIA, and results were correlated with clinical diagnosis. Results In the homogenous pattern noted by IIF, the clustered antigens identified by LIA included dsDNA, Nucleosome, Histone, and Mi-2. In the speckled pattern, the identified antigens were SS-A/Ro52, P0, SS-A/Ro60, SS-B/La, and U1-snRNP. On the other hand, the nucleolar pattern revealed antigens AMA M2, PCNA, and CENP-B. The centromere pattern was mostly associated with CENP-B. The speckled pattern was found to be most commonly associated with systemic lupus erythematosus (SLE). The most common autoantibody found in total ANA profile-positive samples was anti-U1-snRNP followed by anti-SS-A/Ro60 and anti-SS-B/La, and all three were found to be associated with SLE. Conclusions SLE was the most common AD identified in our study samples, with the speckled pattern being the most common pattern on IIF and anti-U1-snRNP being the most common ANA identified by LIA. The fluorescence pattern of IIF predicts the presence of specific antibodies. LIA should be reserved for IIF-positive but dubious cases and whose signs and symptoms are nebulous and do not match the disease dictated by IIF.

A Pilot Study on Blood Components in COVID-19 Affected Subjects: A Correlation to UPR Signalling and ER-Stress.

Abstract: The endoplasmic reticulum (ER) is the site for protein synthesis, its folding and secretion. An intricate set of signalling pathways, called UPR pathways, have been evolved by ER in mammalian cells, to allow the cell to respond the presence of misfolded proteins within the ER. Breaching of these signalling systems by disease oriented accumulation of unfolded proteins may develop cellular stress. The aim of this study is to explore whether COVID-19 infection is responsible for developing this kind of endoplasmic reticulum related stress (ER-stress). ER-stress was evaluated by checking the expression of ER-stress markers e.g. PERK (adapting) and TRAF2 (alarming). ER-stress was correlated to several blood parameters viz. IgG, pro- and anti-inflammatory cytokines, leukocytes, lymphocytes, RBC, haemoglobin and PaO2/FiO2 ratio (ratio of arterial oxygen partial pressure to fractional inspired oxygen) in COVID-19 affected subjects. COVID-19 infection was found to be a state of protein homeostasis (proteostasis) collapse. Changes in IgG levels showed very poor immune response by the infected subjects. At the initial phase of the disease, pro-inflammatory cytokine levels were high and anti-inflammatory cytokines levels were low; though they were partly compromised at later phase of the disease. Total leukocyte concentration increased over the period of time; while percentage of lymphocytes were dropped. No significant changes were observed in cases of RBC counts and haemoglobin (Hb) levels. Both RBC and Hb were maintained at their normal range. In mildly stressed group, PaO2/FiO2 ratio (oxygenation status) was in the higher side of normal range; whereas in other two groups the ratio was in respiratory distress syndrome mode. Virus could induce mild to severe ER-stress, which could be the cause of cellular death and systemic dysfunction introducing fatal consequences. Graphical Abstract: Schematic representation of SARS-CoV-2 infection and related consequences.

Posttreatment Persistence of Monoclonal Protein on Immunofixation Electrophoresis but Absence on Serum Protein Electrophoresis in a Case of Solitary Bone Plasmacytoma.

Laboratory investigations for any suspected case of solitary plasmacytoma of bone include routine biochemical and hematological investigations along with ß2-microglobulin, electrophoresis of serum protein and/or 24-hour urine protein, serum protein immunofixation (IFE), and nephelometric quantification of total immunoglobulin isotype and serum free light chain levels. Bone marrow aspirate and trephine biopsy are mandatory to confirm the absence of clonal plasma cells (for solitary plasmacytoma) or the presence of less than 10% clonal cells (solitary plasmacytoma with minimal bone marrow involvement). Imaging studies such as X-ray, computed tomography (CT), magnetic resonance imaging, and positron emission tomography/CT should be used to complement laboratory tests in diagnosis, staging, and defining the local extent of the plasmacytoma. However, guidelines regarding choice of technique for the detection of M band when monitoring a follow-up case of operated plasmacytoma are still not clear. Through this case study, we try to highlight the role of IFE in a follow-up case of operated solitary plasmacytoma of the bone.

Variation in biochemical parameters in COVID-19 patients admitted at a tertiary care dedicated COVID hospital: A prospective study.

Introduction: COVID-19 infection has a myriad of presentation. Rural India and other developing nations are relatively resource poor, not having access to modern specialized investigations. In this study, we tried to evaluate only biochemical parameters in predicting the severity of the infection. The aim of this study was to find a cost-effective means to predict the clinical course at the time of admission and thereby to reduce mortality and, if possible, morbidity by timely intervention. Materials and Methods: All COVID-19-positive cases admitted at our hospital from March 21 to December 31, 2020, were recruited in this study. The same acted as sham control at recovery. Results: We observed a significant difference in biochemical parameters at the time of admission and discharge, between mild/moderate disease and severe disease. We found slightly deranged liver function tests at admission, which becomes normal at the time of discharge. Urea, C-reactive protein (CRP), procalcitonin, lactate dehydrogenase, and ferritin concentrations in severe/critical patients were significantly higher than that in the mild/moderate group. Receiver operating characteristic curves were plotted to predict the severity on the basis of biochemical parameters independently, of the patients based on these values. Conclusion: We proposed cutoff values of certain biochemical parameters, which will help in judging the severity of the infection at admission. We developed a predictive model with a significant predictive capability for CRP and ferritin values, using normal available biochemical parameters, routinely done in resource-poor centers. Clinicians working in resource-poor situations will be benefitted by having an idea of the severity of the disease. Timely intervention will reduce mortality and severe morbidity.

Résumé Introduction: L'infection au COVID19 a une myriade de présentations. L'Inde rurale et d'autres pays en développement sont relativement pauvres en ressources, non avoir accès aux enquêtes spécialisées modernes. Dans cette étude, nous avons essayé d'évaluer uniquement les paramètres biochimiques pour prédire la gravité de l'infection. Le but de cette étude était de trouver un moyen rentable de prédire l'évolution clinique au moment de l'admission et ainsi de réduire la mortalité et, si possible, la morbidité par une intervention rapide. Matériels et méthodes: Tous les cas positifs au COVID19 admis à notre hospitalisés du 21 mars au 31 décembre 2020, ont été recrutés dans cette étude. La même chose a agi comme un contrôle factice lors de la récupération. Résultats: Nous avons observé une différence significative dans les paramètres biochimiques au moment de l'admission et de la sortie, entre une maladie légère/modérée et une maladie grave. Nous avons trouvé des tests de la fonction hépatique légèrement dérangés à l'admission, qui deviennent normaux au moment de la sortie. Urée, protéine Créactive (CRP, les concentrations de procalcitonine, de lactate déshydrogénase et de ferritine chez les patients sévères/critiques étaient significativement plus élevées que chez les patients légers/modérés groupe. Les courbes caractéristiques de fonctionnement du récepteur ont été tracées pour prédire la gravité sur la base de paramètres biochimiques indépendamment, deles patients en fonction de ces valeurs. Conclusion: Nous avons proposé des valeurs seuils de certains paramètres biochimiques, qui permettront de juger de la gravité de l'infection à l'admission. Nous avons développé un modèle prédictif avec une capacité prédictive significative pour les valeurs de CRP et de ferritine, en utilisant les paramètres biochimiques normaux disponibles, systématiquement effectués dans les centres pauvres en ressources. Les cliniciens travaillant dans des situations où les ressources sont limitées bénéficier d'avoir une idée de la gravité de la maladie. Une intervention rapide réduira la mortalité et la morbidité grave. Mots-clés: COVID19, ferritine, lactate déshydrogénase, urée.

Diagnostic Accuracy of FIB-4 and FIB-5 Scores as Compared to Fibroscan for Assessment of Liver Fibrosis in Patients With Non-Alcoholic Fatty Liver Disease.

Introduction Limited access/exorbitant cost of fibroscan and the associated risks with biopsy to assess fibrosis in non-alcoholic fatty liver disease (NAFLD) patients has made exigent demand of serum-based fibrosis scores to be validated for their accuracy and efficacy. The objective of the study was to compare the accuracy of FIB-4 (fibrosis-4) and FIB-5 (fibrofast) scores to rule out advanced fibrosis in NAFLD patients. Methods A total of 145 patients were categorized as group I with mild/moderate fibrosis (MF) comprising of F0 to F2 and group II with advanced fibrosis (AF) comprising of F3 and F4 based on fibroscan kPa (kilopascal) score. Results Group II had significantly higher alanine transaminase (ALT), aspartate transaminase (AST), haemoglobin % (Hb %), bilirubin and alkaline phosphatase (ALP) values and significantly lower platelet count and albumin as compared to group I. The FIB-4 score was significantly higher in group II [1.8 (1.1 - 4.7)], as compared with group I [0.98 (0.63 - 1.67)], p-value = 0.0001. FIB-5 score of group II [-6.4 (-8.8 - 3.4)] was significantly lower as compared with group I [-4.8 (-6.8 - 2.0)], p-value = 0.003. FIB-4 and FIB-5 had area under receiver operator characteristic (AUROC) curve of 0.712 and 0.655, respectively. FIB-4 at cut-off of <2.02 had a negative predictive value (NPV) of 90.7%. FIB-5 at a cut-off of <-7.11 has an NPV of 94.1% and at a cut-off of <-3.24 had an NPV of 88.9%. Conclusion We concluded that both FIB-4 and FIB-5 can be used to rule out advanced fibrosis in NAFLD patients in a resource-limited and indigent setting as both the scores have NPV greater than 90%.