A pilot study on COVID-19 T-Cell in vaccinated healthcare workers, using Interferon-gamma Release Assay

As is known T-cells play a central role in the immunological response [1]. Nowadays new assays are being developed for the indirect quantification of T-cell memory activity [2]. The aim of this work was to demonstrate Interferongamma Release Assay (IGRA) test could be useful for vaccination monitoring. 23 vaccinated healthcare workers were enrolled in the study after 8 months of the Pfizer BioNTech vaccination. The antibody levels were assessed through Chemiluminescence immunoassay. T cells were indirectly analyzed by an ELISA against INFgamma. Lymphocyte subtyping was evaluated. Statistical analyses were processed. The patients were divided into 3 different groups based on S-RBD and ACE-2 antibody levels: the S-RBD and ACE-2 antibodies were significantly lower in Group 1 than in Group 2 (p<0.001). However, T cells revealed no significant difference between Group 1 and Group 2. Group 3 was the negative control. The results supported the actual role of SARS-CoV-2 T cell, expressed after the vaccine administration and persisting at high concentration over time, despite the antibody levels [3] [4]. Consequently, the new IGRA test was revealed to be an immunological screening that offers information on the protection from SARS-CoV-2 and suggests new strategies for doses administration.

Validation of new Serum Amyloid A chemiluminescence immunoassay with Maglumi 800

Serum Amyloid A (SAA) is an acute-phase protein mainly produced by the liver in response to pro-inflammatory cytokines. SAA is primarily produced by hepatocytes in response to the inflammatory cytokines tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta and IL-6. In healthy individuals, plasma SAA level is within 0.10 mg/L. However, under inflammatory conditions, plasma SAA levels can increase exponentially, even reaching 1000 mg/L or more in some cases. Human SAA expression is upregulated during the acute phase of various viral infections, including cytomegalovirus, herpes simplex virus, measles virus, mumps virus, rubella virus and varicella-zoster virus, and returns to normal during the convalescent phase of infection. Moreover, can be a useful biomarker to predict COVID-19 patient's severity and prognosis.The aim of this study was to evaluate a new chemiluminescence immunoassay for SAA.All serum samples were measured on Maglumi (Snibe platform) and compared with BN ProSpec (Siemens platform), which is used in the routine of the clinical laboratory of the 'Tor Vergata' University Hospital. Analytical precision, correlation coefficient and linearity were assessed. The precision of CLIA system was evaluated by using the commercial normal and high-quality control materials (IQC) recommended by the manufacturer for evaluating precision of SAA. Precision estimation was performed by evaluating triplicate measurements of aliquots of the same samples, performed for a total of five non-consecutive days. Precision data correlated with those declared by the manufacturer. The linearity test was performed using a series of serial dilutions (1/1, 1/2, 1/4, 1/8, 1/16, 1/32, 1/64, 1/128, 1/256 and only diluent) with dilution sample by manufacture. The linearity test showed a correlation coefficient equivalent to 0.997. The results from Snibe platform correlated well with those obtained by Siemens platform with a correlation coefficient of 0.974 (p<0.001). This study demonstrated that the new Snibe SAA test has excellent analytical performance and good reliability and can be used in routine analysis.

1-year monitoring of antibody response after SARSCoV- 2 vaccination in healthcare workers

The global strategy to control coronavirus disease 2019 (COVID 19) was based on the availability of COVID-19 vaccines [1]. Measurement of post-vaccination neutralizing antibodies (Abs) titer, has been shown to be related to protection from SARS-CoV-2 infection [2]. This work aims to improve vaccination data through the evaluation of neutralizing antibodies in triple-dose individuals. To this end, we have conducted a surveillance program focusing at measuring the concentration of IgG Abs against the Receptor Binding Domain (RBD) and neutralizing Abs (NT) anti-SARS-CoV-2 that block the interaction between RBD and the surface receptor cellular angiotensin converting enzyme (ACE2), in the serum of individuals in the vaccination course. The study was conducted on workers from the University of Rome "Tor Vergata" (nTOT=169) who received the Vaxzevria/AstraZeneca vaccine (n=56) and on healthcare workers of the PTV University Hospital who received the Comirnaty/Pfizer-BioNTech vaccine (n=113). Initially for both vaccines two doses were administered: Vaxzevria (12 weeks apart), Comirnaty (2 weeks apart). After the second dose, for the two vaccines has been registered an increase in Abs values both for RBD and NT Abs. As the second dose of the two vaccines has been given at very different time, Pfizer vaccine resulted to response with a higher Abs values earlier in time than Astrazeneca. Moreover, Abs values recorded for those who received Pfizer vaccine are higher up to an order of magnitude. After 6 months from the first dose, the average value Abs titer was 300 BAU/ml and 200 BAU/ml for Pfizer and Astrazeneca respectively. All patients received the Pfizer vaccine as third dose. This last dose gave rise again to an increase of the Abs levels, the average values obtained were 5300 BAU/ml and 3900 BAU/ml for Pfizer and Astrazeneca respectively. As concern NT Abs, we observed a similar pattern to RBD one. After 5 months from the third dose, almost one year from the first dose, antibodies level was over 1000 BAU/ml. Recent work provided Abs cut-off value of immunity against SARSCoV- 2 infection. Values reported range from 200 to 600 BAU/ml [3,4].From this perspective our data have shown a low risk of infection after 1 one year for subjects with a complete vaccine cycle.

Circulating calprotectin as a marker of inflammation and sepsis

Background: Sepsis is an infection-induced syndrome of pathological and biochemical abnormalities believed to be a serious public health problem. The Third International Consensus (Sepsis-3) defined sepsis as 'a life-threatening organ threat caused by a dysregulated host response to infection'. Besides, sepsis patients with concomitant coronavirus disease (COVID-19) have been related to high morbidity and mortality rate. By its identification and characterization, fecal calprotectin (CP) has emerged as a biomarker for intestinal bowel diseases. Recently, its concentration has been documented to increase in serum and plasma samples of patients affected by autoimmune diseases. Since, the biochemical markers used in laboratory diagnosis are often unable to detect the onset of sepsis in a timely manner and given the growing interest in circulating calprotectin (cCP) as a generic marker of inflammation in recent literature, its role has been investigated in emergency room patients requiring hospitalization. Method(s): Patients were divided into four groups: healthy subjects (CTRL);patients hospitalized for causes other than sepsis and negative to SARS-CoV-2 infection (GROUP 1 'NO SEPSIS');septic patients negative to SARS-CoV-2 infection (GROUP 2 SEPSIS) and septic patients positive to SARS-CoV-2 infection (GROUP 3 SEPSICOV). The cCP concentration was determined by a fully automated chemiluminescence immunoassay. Result(s): Circulating calprotectin values detected were significantly higher in groups 1, 2 and 3 (2.08 mug/mL;4.10 mug/mL;2.25 mug/mL), compared to the control group (0.50 mug/ mL) (p <0.0001), and in group 2 'SEPSIS', compared to group 1 'NO SEPSIS' (p<0.0028). The receiving operating characteristic (ROC) curves showed good AUC (area under curve)(0.888;0.945;0.922), sensitivity (80%, 86%;81%) and specificity (99%) values in group 1, 2 and 3, discriminating healthy subjects from patients. To find a cut-off value able to identify sepsis patients, data from group 2 and 3 were combined and compared to group 1 (AUC=0,598;sensitivity=60%;specificity=63%). Conclusion(s): Our results confirmed cCP as a marker of inflammation. Furthermore, the increase in cCP levels in patients with sepsis, suggests its importance as a good marker of sepsis.

Predictive value of Hepcidin level in COVID-19 intensive care unit patients

Coronavirus disease 2019 (COVID-19) caused by the recently uncovered human coronavirus SARS-CoV-2 [1] presents a clinical spectrum that ranges from an asymptomatic condition to critical illness [2]. Patients with critical illness present respiratory failure, septic shock and/ or multi-organ failure induced by the so-defined cytokines storm [3]. Hepcidin modulates cellular iron export to plasma and extracellular fluid through ferroportin, which acts as hepcidin receptor and cellular iron exporter in vertebrates. Hepcidin is not routinely measured in COVID-19 patients, but some preliminary studies showed that high levels of hepcidin are associated with the severity of disease [4] as well as low levels of serum iron are correlated with the severity and mortality of disease and severe hypoxemia in intensive care unit (ICU) patients [5]. The aim of this study was to analyze retrospectively the levels of hepcidin in a group of COVID-19 patients admitted to the intensive care unit (ICU) of the Policlinico Tor Vergata of Rome, Italy. Thirty-eight patients from November 2020 to May 2021 because of pneumonia caused by SARS-CoV-2 were enrolled in the study. Based on the clinical outcome, the patients were assigned to two groups: survivors and non-survivors. A series of laboratory parameters were monitored during the stay of the patients in the ICU and their levels correlated to the clinical outcome. The Hepcidin was determined with Intrinsic Hepcidin IDxTM ELISA kit (Intrinsic Lifesciences, San Diego, CA, USA) that is a competitive immunoenzymatic assay based on a monoclonal antibody (mAb) with high affinity to the Nterminus of hepcidin-25. Of the parameters measured, significant statistical differences in the level of hepcidin, IL-6, LDH, leucocyte count, LNR, NLR, neutrophils level, CRP, and TNF-alpha were observed between the survivors and non-survivors groups. Specifically, a higher level of hepcidin, IL-6, LDH, leucocyte count, LNR, NLR, neutrophils, CRP and TNF-alpha were measured in the non-survivor group compared to the survivor group. In conclusion, Hepcidin can be prognostic of the clinical outcome, moreover, it could be used together with other markers in a predictive algorithm of disease severity.

Autoimmune profile evaluation on vaccinated subjects divided in first, second and booster dose

Background: Coronavirus disease 2019, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), rapidly disseminated worldwide. The clinical research for developing vaccines takes several years but considering the state of emergency, all scientific and technological forces have concentrated towards the formulation of vaccines for the disease containment. In less than one year (December 2020) a first messenger RNA vaccine (Comirnaty, BioNTech/ Pfizer) was authorized. However, the vaccines were administered in phase 4 and the scientific community investigated about eventual side effects on the immune system, discussing the vaccination strategy and possible autoimmune implications. Aim(s): Since the vaccines development process has undergone an unprecedented acceleration, to evaluate a relationship between SARSCoV-2 vaccine administration and impact on the autoimmune level, the determination of circulating immune-complexes (CICs) concentrations, the presence of anti-nuclear antibodies (ANA) and second level tests (dsDNA, ENAscreen and ENAprofile), were studied on vaccinated subjects, after first, second and third dose of Pfizer vaccine. Method(s): The recruited subjects were divided according to anti-SARS-CoV-2 IgG RBD antibodies concentrations in: Group I <10 BAU/ml (N=114);Group II >1000 BAU/ml (N=112);Group III >2500 BAU/ml (N=78). CICs were determined by commercial ELISA kit;ANA by indirect immunefluorescence on Hep-2 cells. Result(s): CICs concentration, did not show significant differences, giving the following median values: Group I =5,44 U/ml;Group II =5,49 U/ ml;Group III =4.29 U/ml.Regarding ANA test: 23.7% of samples (27/114) were positive at 1:80 screening dilution and 4.4% (5/114) at 1:160 clinically relevant dilution, in Group I;15.2% of samples (17/112) were positive at 1:80 screening dilution and 2.7% (3/112) at 1:160 dilution, in Group II;finally 10.2% of samples (8/78) were positive at 1:80 screening dilution and 1.2% (1/78) at 1:160 dilution, in Group III. No specific positivity was found in any group. Conclusion(s): our study did not show significant results variations in the different vaccinated populations examined, suggesting the exclusion of a correlation between vaccine administration and the onset of autoimmune disorders.

SARS-CoV-2 Igg seroprevalence in IBD patients treated with biologics: first vs. second pandemic wave in a prospective study.

OBJECTIVE: In a prospective study, SARS-CoV-2 IgG seroprevalence was assessed during the second pandemic wave (W2) in a cohort of Inflammatory Bowel Disease (IBD) patients using biologics. The secondary aim was to compare, in the same cohort, the frequency of seropositivity and of COVID-19 during the second vs. the first (W1) wave. PATIENTS AND METHODS: From November 2020 to March 2021, SARS-CoV-2 IgG seropositivity and the prevalence of COVID-19 were assessed in a cohort of IBD patients using biologics already studied at W1. INCLUSION CRITERIA: age ≥ 18 years; diagnosis of IBD; follow-up; written consent. EXCLUSION CRITERIA: SARS-CoV-2 vaccination. Risk factors for infection, compatible symptoms, history of infection or COVID-19, nasopharyngeal swab test were recorded. Data were expressed as median [range]. The χ2 test, Student's t-test, logistic regression analysis was used. RESULTS: IBD cohort at W1 and W2 included 85 patients: 45 CD (52.9%), 40 UC (47.1%). When comparing the same 85 patients at W2 vs. W1, a higher SARS-CoV-2 seroprevalence at W2 was at the limit of the statistical significance (9.4% vs. 2.3%; p=0.05). The prevalence of COVID-19 at W2 vs. W1 was 3.5% (3/85) vs. 0% (0/85) (p=0.08). Contacts with COVID-19 patients and symptoms compatible with COVID-19 were more frequent at W2 vs. W1 (18.8 % vs. 0%; p=0.0001; 34.1% vs. 15.3%; p=0.004). At W2, history of contacts and new onset diarrhea were more frequent in seropositive patients [4/8 (50%) vs. 12/77 (15.6%); p=0.01 and 4/8 (50%) vs. 2/77 (2.6%); p=0.0001]. At W2, the risk factors for seropositivity included cough, fever, new onset diarrhea, rhinitis, arthromyalgia, dysgeusia/anosmia at univariate (p<0.05), but not at multivariate analysis. History of contacts was the only risk factor for seropositivity at univariate (p=0.03), but not at multivariate analysis (p=0.1). CONCLUSIONS: During W2, characterized by a high viral spread, IBD and biologics appeared not to increase the prevalence of SARS-CoV-2 infection or COVID-19 disease. New onset diarrhea mimicking IBD relapse may be observed in patients with SARS-CoV-2 infection.

Lessons learned from the COVID-19 pandemic: emphasizing the emerging role and perspectives from artificial intelligence, mobile health, and digital laboratory medicine

SARS-CoV-2, the new coronavirus causing COVID-19, is one of the most contagious disease of past decades. COVID-19 is different only in that everyone is encountering it for the first time during this pandemic. The world has gone from complete ignorance to a blitz of details in a matter of months. The foremost challenge that the scientific community faces is to understand the growth and transmission capability of the virus. As the world grapples with the global pandemic, people are spending more time than ever before living and working in the digital milieu, and the adoption of Artificial Intelligence (AI) is propelled to an unprecedented level especially as AI has already proven to play an important role in counteracting COVID-19. AI and Data Science are rapidly becoming important tools in clinical research, precision medicine, biomedical discovery and medical diagnostics. Machine learning (ML) and their subsets, such as deep learning, are also referred to as cognitive computing due to their foundational basis and relationship to cognition. To date, AI based techniques are helping epidemiologists in projecting the spread of virus, contact tracing, early detection, monitoring, social distancing, compiling data and training of healthcare workers. Beside AI, the use of telemedicine, mobile health or mHealth and the Internet of Things (IOT) is also emerging. These techniques have proven to be powerful tools in fighting against the pandemic because they provide strong support in pandemic prevention and control. The present study highlights applications and evaluations of these technologies, practices, and health delivery services as well as regulatory and ethical challenges regarding AI/ML-based medical products. © 2021 International Federation of Clinical Chemistry and Laboratory Medicine. All rights reserved.

Impact of SARS-CoV-2 on dentistry: a review of literature.

OBJECTIVE: SARS-CoV-2 is a new Coronavirus identified as the cause of Coronavirus disease in 2019 (COVID-19). The epidemic spread in China and beyond its borders, involving 114 countries with more than 5 million dead. On March 11, the WHO declared the spread of SARS-CoV-2 to be a pandemic and encouraged nations to adopt harsh restrictive measures. Therefore, patients more and more often turn to dental offices only for emergencies. Healthcare professionals, including dentists, are at high infectious risk. In fact, the closeness to the oral cavity and nasopharynx and the use of drills or ultrasonic devices that cause aerosol release, make dental professions at high risk of bacterial and viral infections. The way patients are treated has changed. In fact, it should be mandatory to carry out a pre-treatment telephone triage and the use of mouthwashes to reduce bacterial load. In the current pandemic, it is necessary to adopt specific safety protocols that can protect dental operators as well as limit the spread of the virus. The purpose of this review is to present an overview on ways to reduce the risk of SARS-CoV-2 contagion in dentistry by focusing on the immediate situation as well as by looking towards the future. MATERIALS AND METHODS: To reach the review purpose, we selected a series of studies using keywords "COVID-19" OR "SARS-CoV-2" in association with "dentistry" AND "safety protocols" AND "healthcare procedures" AND "individual protection dispositive" AND "air transmission" AND "droplet". We selected papers exclusively in English language, up to 1st January 2022. RESULTS: During future phases of the pandemic, everywhere in the World, it is necessary to impose all dentistry team both a serological screening and the vaccination, as already established for all health staff in Italy. CONCLUSIONS: For own safety, it is an important for the whole dentistry category constantly update the devices and the protocols adopted, as well as monitoring the real infectious threats, which may occur.

The great value of clinical laboratories in the pandemic process

Laboratory Medicine has been showing its great Value during Covid-19 pandemic in the whole care process: diagnosis, screening, follow up and outcome of the disease. The diagnostic tools to date are able to dose viral RNA, and antibodies against viral proteins or viral proteins themselves. Even if the conventional RT-PCR has been the most widely used method, the greatest innovation in molecular diagnosis was reported by the so called CRISPR Community. The first serological tests to be introduced were rapid serological tests with direct reading (first generation) or with fluorescence reading (2nd generation) and microfluidic with fluorescent reading (3rd generation). Then conventional CLIA method have been introduced with better sensitivity and more recently a new kind of serological assay has been proposed able to detect anti SARS-Cov-2 serum antibodies throughout a competitive streptoavidin/biotin assay which utilize RBD as coated antigen and ACE-2 labelled with biotin as competitor molecule for serum antibodies. Until few months ago the harmonization between many methods produced by many manufacturers was impossible. Recently the WHO produced an international standard (pool of sera-total antibodies) able to permit a sort of harmonization. To date NGS return to be widely utilized in the effort to intercept new variants. Finally, pathophysiology and natural course of Post-acute Sequelae of COVID-19 (PASC) also called Long Covidis unclear, meriting further studies. Also in this almost still unknown field, Clinical Laboratory could contribute in diagnosis and monitoring.

Evaluation of ECLIA antigen detection tests as screening methods for COVID-19 in comparison with molecular analysis

In the ongoing COVID-19 pandemic, rapid diagnostic testing for SARS-CoV-2 is necessary to limit virus spread. Different diagnostic rapid tests have been developed as rapid and helpful tools for diagnosis of COVID-19, based on virus proteins detection in respiratory samples.This study aims to evaluate the performances of the Elecsys SARS-CoV-2 Antigen test, in comparison to rRT-PCR, the gold standard.Molecular analysis was carried on 110 swabs from Lifebrain laboratory. According to rRT-PCR, 76 samples were positive, 34 were negative. Initially, the sensitivity and specificity were 85% and 100%, respectively. However, since most of the discordant cases had cycle threshold (Ct) values > 28, it was assumed a new measure to evaluate sensitivity and specificity. At this point, samples with Ct values <28 were selected and a sensitivity of 94% was achieved. The level of agreement between the two tests was 89,1% with η value of 0,77 for total data and 95,9% with η value of 0,95 for samples with <28 Ct.The Antigen test is a well performed tool, timely and effortless, in presence of high viral loads. The comparison data validated the method as a proper approach for rapid screening of patients with high SARSCoV-2 viral load. Also, a double test for confirmatory analysis on the same swab could increase the overall lab-workflow, but the rate of sensitivity is still highly Ctdependent.

Evaluation of SARS-CoV-2 S-RBD IgG and NAb antibody levels in vaccinated subjects

BACKGROUND: Covid 19 disease represents the largest public health emergency.Following the spread of Covid vaccines, it has become of central importance for laboratories to assess immunity, protection against SARS-CoV-2 and whether booster shots will be needed.Aims of the study are:Detection of antibodies following SARS-CoV-2 vaccination;Monitoring of anti-Spike SARS-CoV-2 antibody levels (S-RBD) induced by vaccination;Monitoring of neutralizing SARS-CoV-2 antibody levels (NAb) induced by vaccination;Identifying antibody levels of previously infected subjects;Correlation between antibody levels and type of vaccine used;METHODS: A total of 70 workers from the University 'Tor Vergata' in Rome and from the University hospital, were monitored during their vaccination program (Astra Zeneca and Pfizer-BioNTech vaccines). Serum samples collected at different time points 10, 20, 35, 50, 80, 120, 150 days after the first and second dose of vaccine. A chemiluminescent assay for the quantitative determination of SARS-CoV-2 S-RBD IgG and NAb was used, performed on the fully automated Mindray CL 1200i analytical system.RESULTS: The antibody concentrations detected in the two groups of workers after the first dose made us able to distinguish them into three different groups: subjects previously naturally infected, with higher antibody production;uninfected with the lowest antibody concentration values and a group with antibody values in the middle between the two, probably workers with asymptomatic infection. The amount of S-RBD and NAb antibodies in vaccinated subjects with pre-existing immunity is almost as twice as high than in naive vaccinated subjects at the same time points. Both vaccinations, although with differentiated antibody concentrations, reach a peak about 30 days after the first dose and then decrease up to 150 days, stopping at a steady state of around 150 -200 BAU/ml. CONCLUSIONS: These data highlights: the importance of serologic testing before vaccination in order to distinguish previously infected asymptomatic persons thus avoiding possible side effects such as the development of antibody-dependent enhancement (ADE);the importance of completing the two doses recommended for noninfected subjects in order to achieve strong levels of immunity.

The assessment of high sensitivity cardiac troponin in patients with COVID-19: A multicenter study

Background: Recent studies have shown that patients diagnosed with coronarivus disease 2019 (COVID-19) and also with previous cardiovascular diseases have a higher mortality due to worsening heart disease. At the same time, patients without previous cardiovascular disease may also have cardiac complications. The aim of this multicenter study was to assess high sensitivity cardiac troponin T (hs-cTnT) in patients with COVID-19 and to evaluate the incidence of myocardial injury. Methods: In this multicenter study we enrolled 543 patients, 57.8% males, median age 63 years (range 18-99) from three selected hospitals: University Hospital Tor Vergata in Rome, Fondazione IRCCS Ca 'Granda Ospedale Maggiore Policlinico, in Milan, S Chiara Hospital in Trento. We measured hs-cTnT in all patients to assess myocardial injury and correlations with patient's age, symptoms and disease course. Results: The data showed that, among the 543 patients studied, 257 patients (47.3%) had hs-cTnT values higher than the upper reference limit (URL) of 14 ng/L. Patients with high hscTnT had more frequently fever (p < 0.01) and respiratory symptoms (p < 0.01), compared to the group with hscTnT values below URL. The results showed also that patients with hs-cTnT above URL had a higher frequency of previous cardiovascular disease (p < 0.01) as well as of hypertension (p < 0.01). Instead, among 231 patients with no previous cardiovascular disease, 81 (31.5%) had hs-cTnT values above the URL. Finally. the majority of the patients with high hs-cTnT were admitted to the intensive care unit (p < 0.01). Conclusion: Our data suggest the assessment of high sensitivity cardiac troponin in patients with COVID-19 for early diagnosis of cardiac involvement.

Leucocyte differential and morphometric parameters with mindray bc-6800 plus: A possible predictive tool to diagnose sepsis and sars-cov-2 infections

Background: Sepsis is an infectious disease (the etiology can be viral or bacterial) with hight mortality, threatening human health. Clinicians need to diagnose the patient's infection in time and look for pathogens in order to develop an effective treatment plan;therefore, a quickly and early screen to diagnose sepsis has become an urgent problem in clinical laboratories. Different inflammatory factors are used to diagnose the sepsis;CRP, IL-6, PCT, ADM, lactate, D-dimer etc., but they also have limitations such as insufficient sensitivity and specificity and requiring additional examination cost. The aim of this study is to use leucocyte counts (neutrophils and monocytes that are activated from pathogenic virus or bacteria) and others morphological change with Mindray BC-6800-plus platform to diagnose sepsis early, quickly, conveniently and at low cost. Methods: A total 957 EDTA-k2 anticoagulant venous whole blood samples were collected: 70 control patients (blood donors) with a normal complete count blood and negative VES, and 887 samples hospitalized at the emergency department with symptoms attributable to sepsis with PCT request. All data was divided in 4 groups: control group, group where sepsis cannot be confirmed, group with confirmed sepsis diagnosis and a group with sepsis from SARS-CoV-2 infection. Morphometric and numeric parameters are reported with Mindray BC-6800 plus: blood count like positional parameters X, Y, Z of neutrophils, lymphocites and monocytes, PLT, NLR (neutrophil lymphocyte ratio) and IMG (index of immature granulocytes). For statistical analysis was used Shapiro Wilk test for distribution analysis and the non parametric Kruskal Wallis test to evaluate significative differences among the groups (p< 0.05) and also examined ROC curve analysis. Results: There is a statistically significant difference between control group and sepsis group for haematological parameters: positional parameters (Neu X, Y, Z;Mon X, Y, Z and Lym X, Y, Z), IMG, NLR, PLT. The roc curves highlight acceptable sensitivity and specificity values for some haematological parameters and suggest a possible cut-off. Conclusions: The BC-6800 plus can help the diagnosis of sepsis upon the admission to the emergency department using some morphological positional parameters.

Low testosterone levels and high estradiol to testosterone ratio are associated with hyperinflammatory state and mortality in hospitalized men with COVID-19.

OBJECTIVE: Evidence supports a sex disparity in clinical outcomes of COVID-19 patients, with men exhibiting higher mortality rates compared to women. We aimed to test the correlation between serum levels of sex hormones [total testosterone, estradiol (E2), estradiol to testosterone (E2/T) ratio, progesterone), prolactin and 25-hydroxyvitamin D [25(OH)D] and markers of inflammation, coagulation and sepsis at admission in hospitalized men with COVID-19. PATIENTS AND METHODS: We conducted an exploratory retrospective study including symptomatic men with confirmed SARS-CoV-2 infection who were consecutively admitted to our Institution between April 1 and May 31, 2020. RESULTS: Patients were divided into survivors (n=20) and non-survivors (n=39). As compared to survivors, non-survivors showed significantly higher median neutrophil-to-lymphocyte ratio (NLR) values, D-dimer and procalcitonin (PCT) levels, along with significantly lower median 25(OH)D levels and total testosterone levels. Non-survivors exhibited significantly higher median values of E2/T ratio (a marker of aromatase activity). Spearman's correlation analysis revealed that total testosterone levels were significantly and inversely correlated with NLR, high-sensitivity C-reactive protein (hsCRP), interleukin-6, D-dimer and PCT. Conversely, E2/T ratio values were significantly and positively correlated with the aforementioned markers and with white blood cell (WBC) count. In a multivariate analysis performed by a logistic regression model after adjusting for major confounders (age, body mass index, hypertension and cardiovascular disease, diabetes mellitus and malignancy), total testosterone levels were significantly and inversely associated with risk of COVID-19-related in-hospital mortality. CONCLUSIONS: Low total testosterone levels and elevated E2/T ratio values at admission are associated with hyperinflammatory state in hospitalized men with COVID-19. Low total testosterone levels at admission represent an independent risk factor for in-hospital mortality in such patients. Therefore, total testosterone and E2/T ratio may serve as prognostic markers of disease severity in this population.

Ad interim SIBioC recommendations for serological assessment of SARS-CoV-2 infection

The recent pandemic outbreak caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and associated with the pathology called COVID-19 (coronavirus disease 2019), has now become one of the most strenuous health care challenges since the emergence of the three pandemics caused by influenza viruses during the past century. Throughout the clinical decision-making of COVID-19, laboratory tests are essential for supporting the screening, diagnosis, prognostication and therapeutic monitoring of this severe infectious disease. Serological testing, that reflects the humoral immune response developing after interaction between the host and the virus (or its components), enables to garner a vast array of clinical information which can be especially used in seroprevalence or seroconversion studies. To this end, the Task Force on COVID-19 of the Italian Society of Clinical Biochemistry and Clinical Molecular Biology (SIBioC) has endorsed a series of technical, practical and clinical ad interim recommendations, aimed at facilitating and optimizing the introduction, clinical usage and governance of SARS-CoV-2 serological immunoassays in routine practice.

COVID-19: Serology, vaccines and variants

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has prompted the scientific community and the pharmaceutical companies to put the maximum efforts for developing vaccines able to contain the spread of SARS-CoV-2. Serology tests were produced very soon with different sensitivities and specificities depending from the methods and technologies utilised. Presently, many vaccines have been developed and authorized for use in human beings in different Countries. All of them are based on a version of the spike glycoprotein characterized at the beginning of the pandemic. However, they differ by their level of efficacy against COVID-19. SARS-COV-2 as other RNA viruses mutates continuously. Genome sequencing analysis showed a nucleotide substitution rate of about 1 × 10-3 substitutions per year that lead to the emergence of variants through point mutations, insertions, deletions and recombination. There is concern about the ability of the current vaccines to protect against the emerging viral variants. Mutations in the spike glycoprotein may affect transmission dynamics and the risk of immune escape. In this review, we address the different technological platforms in use for developing COVID-19 vaccines, the impact of emerging viral variants on virus transmission, hospitalization, and response to current vaccines as well as rare but important adverse reactions to them. Finally, different methods for measuring the antibody response to vaccine are reported including the importance of using the WHO international Standard for creating a common language for reporting data.

Serum Amyloid A Protein as a useful biomarker to predict COVID-19 patients severity and prognosis

Coronavirus Disease 2019 (COVID-19) can present with different grades of severity from mild to critical. Evaluation of biomarkers predicting severity is crucial to identify patients at high risk of disease progression and poor prognosis. Serum Amyloid A (SAA) is an acute-phase protein mainly produced by the liver in response to pro-inflammatory cytokines. In this study, we investigated SAA levels at admission (T1) and after 15 days (T2) of hospitalization in two groups of patients: survivors and non-survivors. At T1, the non-survivors showed higher SAA level than survivors (74 mg/dL vs 48.75 mg/dL). At T2, the survivor group value decreased to 6.55 mg/dL, the non-survivor group still showed high levels (51.1 mg/dL). The SAA level in control group was 0.35 mg/dL. Furthermore, a cut-off value of 63 mg/dL able to discriminate survivors from non-survivors was established by ROC curve analysis at T1. At T2, the cut-off decreased to 30.9 mg/dL. A similar decreasing trend was observed for D-Dimer, hsCRP, IL-6 and procalcitonin levels. The results of this retrospective study suggest that SAA is a good marker of COVID-19 disease alone and/or in combination with other inflammatory biomarkers. Identification of reliable prognostic analytes is of great clinical relevance, as it would improve patient management besides being costs saving.

Persistence of SARS-CoV-2 Viral RNA in Nasopharyngeal Swabs after Death: An Observational Study

The aim of this study was to investigate the persistence of SARS-CoV-2 in post-mortem swabs of subjects who died from SARS-CoV-2 infection. The presence of the virus was evaluated post-mortem from airways of 27 SARS-CoV-2 positive patients at three different time points (T1 2 h;T2 12 h;T3 24 h) by real-time PCR. Detection of antibodies to SARS-CoV-2 was performed by Maglumi 2019-nCoV IgM/IgG chemiluminescence assay. SARS-CoV-2 viral RNA was still detectable in 70.3% of cases within 2 h after death and in 66,6% of cases up to 24 h after death. Our data showed an increase of the viral load in 78,6% of positive individuals 24 h post-mortem (T3) in comparison to that evaluated 2 h after death (T1). Noteworthy, we detected a positive T3 post-mortem swab (24 h after death) from 4 subjects who were negative at T1 (2 h after death). The results of our study may have an important value in the management of deceased subjects not only with a suspected or confirmed diagnosis of SARS-CoV-2, but also for unspecified causes and in the absence of clinical documentation or medical assistance.