ABSTRACT
Introduction: SARS-CoV-2 infection has plagued the world for the past two years, during which time it has become necessary to use screening tests to prevent the spread of the virus, especially among high-risk patients. The gold standard for the detection of viral RNA is real-time PCR (RT-PCR). This technique requires special care when handling samples, as well as being time-consuming and costly. To meet the need for mass screening, antigenic tests were developed and continuously improved over the years. In our study, we examined the performance of Lumipulse G SARS-CoV-2 Ag (Fujirebio, Japan), a quantitative and automated chemiluminescence enzymeimmunoassay- based antigen test. Material(s) and Method(s): Our study includes 160 subjects (median age 38 years, interquartile range (IQR) 24-58 years;43% females) screened for SARS-COV-2 at the Service of Laboratory Medicine of Pederzoli Hospital (Peschiera del Garda, Verona, Italy), between August 16 and September 15, 2021. A nasopharyngeal swab was collected for each subject and analyzed at the same time by antigen test Fujirebio Lumipulse G SARS-CoV-2 Ag and by molecular test performed by Altona Diagnostics RealStar SARSCoV- 2 RT-PCR Kit (Altona Diagnostics GmbH, Hamburg, Germany). Result(s): A significant Spearman's correlation was found between values of Fujirebio Lumipulse G SARS-CoV-2 Ag and measurable Ct values of SARSCoV- 2 of both the S (r= -0.94;p<0.001) and E genes (r= -0.95;p<0.001). The sensitivity and specificity at 1.0 pg/ mL (cut-off already used by other authors to discriminate samples positive for SARS-CoV-2) were 0.71 and 1.00. We also used a locally calculated threshold of 0.60 pg/mL (sensitivity 0.88;specificity 0.75). The exclusion of samples tested positive at molecular testing with Ct values comprised between 25-37 enabled the attainment of better diagnostic performance on the 103 residual samples using the 0.60 pg/mL cut-off. Conclusion(s): the results of our study confirm the good performance of Fujirebio Lumipulse G SARS-CoV-2 Ag (considering cutoff 1.0 pg/mL), especially in samples with high viral load (i.e., Ct value <25), which has proved even better using our locally-calculated cut-off (i.e., 0.60 pg/mL).
ABSTRACT
Introduction: In a historical period like the one we are experiencing now, the perspective that we will have to coexist with SARS-CoV-2 for a long time is effective. Hence predicting and monitoring the effectiveness of the vaccine response is a task that has become essential for laboratory medicine. Being secreted by the mucous membranes, de facto IgA constitutes the first barrier against Coronavirus. In this study, we focus on describing the trend of IgA neutralizing antibodies in a court of hospital employees who received a booster dose of vaccine, after completing the primary vaccination cycle. Material(s) and Method(s): The study population consisted of 73 healthcare workers (median age 44 years, 54.8% women) of the Pederzoli Hospital (Peschiera del Garda, Verona, Italy), receiving a booster dose of Pfizer COVID-19 vaccine BNT162b2 (Comirnaty). Serum samples were collected before receiving the booster dose (baseline), one month (T1) and three months (T2) afterward. We then measured anti-spike S1 subunit IgA (ELISA, Euroimmun). Results were presented as the median and interquartile range (IQR) or as a ratio with baseline anti-SARS-CoV-2 antibodies value. Result(s): The median anti-spike S1 subunit IgA titer of our population at baseline was a 1,28 ratio (IQR 0.73-2.21). One month after the booster dose a 4.5-fold increase from baseline occurred (median 5.86 ratio;IQR 4.82-6.22;p<0.001). Three months after vaccination we observed a 16.7% decline of antibody titer from T1 (median 4.88 ratio;IQR 3.45-5.23;p<0.001). 28 subjects, seronegative at baseline had an 8.5-fold increase at 1 month, compared to 45 seropositive subjects at baseline who had only 2.93-fold increase. The decrease at T2 was significantly higher in seronegative group than in seropositive group, respectively 30% and 16,8%. Moreover, only two baseline seronegative subjects out of the entire study population resulted seronegative again at T2. Conclusion(s): Our study shows that there is a substantial increase in antibody titer after booster dose in the whole study population. Considering that 97.3% of them remained seropositive at T2, our results strengthen the importance of adhering to a vaccination campaign in order to maintain an adequate antibody titer and limit the spread of SARS-CoV-2.
ABSTRACT
Introduction: In March 2020, WHO, after assessing the global spread of SARS-CoV-2 infection, declared the outbreak of COVID-19 a pandemic. Several vaccines have been developed during these years, but some studies have shown that their efficacy decreases significantly over time, so a booster dose was required. In November 2021, the administration of the third dose (booster) of the SARS-CoV-2 vaccine was made mandatory. With our study, we want to describe the antibody trend up to 6 months after the booster dose. Material(s) and Method(s): The study population consisted of 255 healthcare workers (median age 47 years, 66,3% women) of the Pederzoli Hospital (Peschiera del Garda, Verona, Italy), receiving a booster dose of Pfizer COVID-19 vaccine BNT162b2 (Comirnaty). Serum samples were collected before the booster dose (baseline), one month (T1), and six months (T2) afterwards. The serum levels of anti-SARS-CoV-2 spike trimeric IgG were assayed with DiaSorin Trimeric spike IgG (DiaSorin, Saluggia, Italy). Results were presented as the median and interquartile range (IQR) or as a ratio with baseline anti-SARS-CoV-2 antibodies value. Result(s): In our population at T1 (median 7730 BAU/mL;IQR 4462,5-12350) we observed an increase (32,6-folds) of anti-SARS-CoV-2 spike trimeric IgG compared to baseline (median 237 BAU/mL;IQR 129,5-416). At T2 (median 3720 BAU/mL;IQR 1502,5-9495) there was a 51,9% decrease from T1. During the observation period (after the booster dose), 111 subjects resulted positive for a nasopharyngeal swab. This subgroup showed a 16.6% increase in antibody titer between T1 and T2, while the non-infected subjects had a significant decrease (73.2%). Furthermore, in subjects aged 50 years or older, the antibody titer dropped by 60.8% from T1 to T2 compared with the titer of subjects <50 years which decreased by 42.4%. Conclusion(s): the booster dose of Pfizer COVID-19 vaccine BNT162b2 triggers an excellent immune response in all subjects included in this study. However, the serum levels of anti-SARS-CoV-2 antibodies gradually lower six months after administration, but they still remain discreetly high. Since the decrease is more pronounced in elderly subjects, the assumption to carry out a second booster dose, at least in the most fragile subjects, becomes more concrete.
ABSTRACT
Background: we report here data on humoral immune response post-BNT162b2 primary vaccination and booster in pre-vaccination baseline severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) seronegative and seropositive subjects. Method(s): the study population consisted in 51 baseline SARS-CoV-2 seronegative and 11 baseline SARS-CoV-2 seropositive subjects, who underwent primary mRNA-based BNT162b2 vaccination (two doses) followed by homologous booster administration (third dose). Venous blood was sequentially collected up to 1 months after vaccine booster administration, and humoral response was monitored by measuring anti-SARS-CoV-2 spike trimeric IgG antibodies. Result(s): the humoral response after the three doses of BNT162b2 displayed an overlapping trend in the two groups, although the baseline and post-primary vaccination concentration of anti-SARS-CoV-2 spike trimeric IgG were constantly higher in baseline SARS-CoV-2 seropositive than in baseline SARS-CoV-2 seronegative subjects (all p<0.001). Unlike before vaccine booster administration, the levels of anti-SARS-CoV-2 spike trimeric IgG, 1 month after receiving the third BNT162b2 dose were not significantly different between pre-vaccination baseline SARS-CoV-2 seropositive and seronegative subjects (7 430 versus 9 020 kBAU/L;p=0.232). In both cohorts, all recipients of vaccine booster displayed antibodies levels >264 kBAU/L. Conclusion(s): the results of this study demonstrate that although baseline SARS-CoV-2 seropositive subjects have magnified humoral response to primary BNT162b2 vaccination, vaccine booster generates anti-SARS-CoV-2 spike trimeric IgG values not different from those found in baseline SARS-CoV-2 seronegative subjects. Thus, this study provides evidence that a prior SARS-CoV-2 infection does not mitigate the need for additional vaccine boosters. Copyright © 2022 Biomedia. All rights reserved.
ABSTRACT
Introduction: COVID-19 vaccination campaign at Pederzoli Hospital (Veneto, Italy) started in January 2021 using the two approved m-RNA vaccines. The vaccination schedule included 2 doses and one booster. Objective(s): The Pharmacy unit created a digital form in order to collect information on the incidence of the adverse events after the anti-covid vaccination. Method(s): The Pharmacy Unit created a simplified digital form including a set of questions about the occurrence of adverse events. To improve the adherence to this pharmacovigilance program, the digital form was uploaded on the hospital website and all the employees received a remainder about this vaccine adverse event reporting system via e-email regularly. The data collected from the electronic spreadsheet were analyzed by the Health Department in order to produce a report in the dedicated national website (Vigifarmaco). Result(s): From January 2021 to April 2022 1076 employees received the first dose, 965 received the second dose, and 1019 received the third dose. 323 adverse events reporting forms have been collected after the first dose (30% of those who received it). 291 forms (30%) were collected after the second dose. 12 forms (1.2%) were collected after the third dose. The most reported adverse events after the first dose were: local pain (38% of reported adverse events), muscle pain 15%;asthenia 12%, headache 11%. Moreover, 6 employees reported fever above 38.5degreeC and 19 employees reported fever between 37.5 degreeC and 38.5degreeC. The most reported adverse events after the second dose were: local pain 19%, muscle pain 18%;asthenia 15%, headache 13%. An employee reported a syncope. In addition,35 reported fever above 38.5 degree C and in 70 fever between 37.5degreeC and 38.5 degreeC. After the third dose, 10 reported headache, 9 local pain, and 8 asthenia and muscle aches. Only 2 employees reported fever above 38.5degreeC and 5 fever between 37.5degreeC and 38.5degreeC. Conclusion(s): The active role of the Hospital Pharmacy Unit during the vaccination campaign of the health care workers of our hospital permitted to collect data regarding the side effects of these new vaccines and nonetheless made the health care workers about more aware about the importance of vaccine adverse event reporting system. The use of this digital form supported the pharmacovigilance program creating a useful and ''user friendly'' tool which may be tailored to new future projects.
ABSTRACT
Introduction: COVID-19 vaccination campaign at Pederzoli Hospital (Veneto, Italy) started in January 2021 using the two approved m-RNA vaccines. The vaccination schedule included 2 doses and one booster. Objective: The Pharmacy unit created a digital form in order to collect information on the incidence of the adverse events after the anti-covid vaccination. Methods: The Pharmacy Unit created a simplified digital form including a set of questions about the occurrence of adverse events. To improve the adherence to this pharmacovigilance program, the digital form was uploaded on the hospital website and all the employees received a remainder about this vaccine adverse event reporting system via e-email regularly. The data collected from the electronic spreadsheet were analyzed by the Health Department in order to produce a report in the dedicated national website (Vigifarmaco). Results: From January 2021 to April 2022 1076 employees received the first dose, 965 received the second dose, and 1019 received the third dose. 323 adverse events reporting forms have been collected after the first dose (30% of those who received it). 291 forms (30%) were collected after the second dose. 12 forms (1.2%) were collected after the third dose. The most reported adverse events after the first dose were: local pain (38% of reported adverse events), muscle pain 15%;asthenia 12%, headache 11%. Moreover, 6 employees reported fever above 38.5 °C and 19 employees reported fever between 37.5 °C and 38.5 °C. The most reported adverse events after the second dose were: local pain 19%, muscle pain 18%;asthenia 15%, headache 13%. An employee reported a syncope. In addition,35 reported fever above 38.5 ° C and in 70 fever between 37.5 °C and 38.5 °C. After the third dose, 10 reported headache, 9 local pain, and 8 asthenia and muscle aches. Only 2 employees reported fever above 38.5 °C and 5 fever between 37.5 °C and 38.5 °C. Conclusion: The active role of the Hospital Pharmacy Unit during the vaccination campaign of the health care workers of our hospital permitted to collect data regarding the side effects of these new vaccines and nonetheless made the health care workers about more aware about the importance of vaccine adverse event reporting system. The use of this digital form supported the pharmacovigilance program creating a useful and "user friendly" tool which may be tailored to new future projects.
ABSTRACT
Introduction: The monocyte distribution width (MDW) reflect volume variation of circulating inflammatory monocytes, cells with high plasticity capable to react against viral infections. Since a few studies have been published on the correlation between coronavirus disease 2019 (COVID-19) and MDW, we further explored this association during the second and third wave of COVID-19 infection in Italy.Materials and Methods: A retrospective study was conducted in two groups of COVID-19 patients, 126 testing positive from November to December 2020 (mean age, 71±15;41 women, 32.53%) and 59 from March to April 2021 (mean age, 67±17;25 women, 42.37%). A third group was also included, composed of 123 non-COVID-19 hospitalized patients (mean age, 62±22, ;62 women, 50.41%). All these patients were hospitalized at the Pederzoli Hospital (Peschiera del Garda, Verona, Italy). UniCel DxH800 Hematology Analyzer (Beckman Coulter Inc., CA, USA) was used to analyze whole blood venous samples previously collected in K2-EDTA, according to routine methods for routine blood cell count (thus including MDW). Results were expressed as median and interquartile range (IQR).Results: MDW values were significantly higher in the entire cohort of COVID-19 patients (median and IQR, 21.8 and 4.4 fL;p<0.001), as well as in the first-wave COVID-19 positive group (median and IQR, 22.0 and 4.1 fL;p<0.001) and in the secondwave COVID-19 positive group (median and IQR 21.5 and 4.1 fL;p<0.001) compared to the control cohort (median and IQR, 17.8 and 2.7 fL). No significant differences in MDW were observed between the two COVID-19 positive groups (p=0.251).Conclusion: Monocytes are cells of innate immunity strongly involved in the pathogenesis of COVID-19 and their size variation, as result of infection and/or activation, is reflected by MDW changes. Overall, our data show that MDW is higher in COVID-19, thus mirroring the hyper-inflammatory condition triggered by SARS-CoV-2 infection. Routine assessment of MDW could hence be useful for diagnosing and monitoring patients with SARS-CoV-2 infection.
ABSTRACT
Introduction: The neutrophil-to-lymphocyte (NLR) is a readily available biomarker reflecting systemic inflammation, derived from a ratio of absolute blood neutrophil and lymphocyte counts. Since coronavirus disease 2019 (COVID-19) requires the identification of early and efficient diagnostic and prognostic biomarkers, we compared the NLR index between subjects with or without positive COVID-19 molecular swab.Materials and Methods: A retrospective study was conducted in two groups (CoV1 and CoV2) of COVID-19 patients, 126 testing positive from November to December 2020 (CoV1;mean age, 71±16;32.5% women) and 59 testing positive from March to April 2021 (CoV2;mean age, 68±18;42.4% women). A third group (CoN, controls) included 123 non-COVID-19 ostensibly healthy subjects (mean age, 63±22;50.4% women). All patients were hospitalized at Pederzoli Hospital (Peschiera d/G, VR, Italy). Whole blood venous samples previously collected in K2-EDTA were analyzed with UniCel DxH800 Hematology Analyzer (Beckman Coulter Inc., CA, USA). Results were expressed as median and interquartile range (IQR).Results: Subjects in group CoV1 had significantly higher NLR values (median and IQR, 6.33 and 9.38;p=0.029) compared to subjects testing negative for COVID-19 (median and IQR, 4.10 and 6.44), whilst no significant difference was found between NLR values of group CoV2 (median and IQR, 4.90 and 5.54;p=0.414) and the control cohort. Accordingly, NLR values in group CoV1 were significantly higher than in CoV2 (p=0.031).Conclusion: In this study, NLR was found significantly higher in COVID-19 patients during the November-December 2020 outbreak, but not during the following March to April 2021 wave. We hypothesize that this may be due to the more pronounced inflammatory state in the former period, which was also associated with a much higher hospitalization and death rate in our region.
ABSTRACT
Background: This observational retrospective study was aimed at evaluating the clinical performance of the novel microfluidic fluorescence immunoassay FREND COVID-19 Ag test in a population of unselected individuals undergoing routine SARS-CoV-2 (severe acute respiratory coronavirus 2) testing. Methods: The study population consisted of a series of outpatients referred to the Service of Laboratory Medicine of Pederzoli Hospital (Peschiera del Garda, Verona, Italy) between April 12 and 30, 2021, for SARS-CoV-2 testing for being either symptomatic or having had close contact with one or more COVID-19 cases. A routine nasopharyngeal sample was collected at hospital admission and analyzed with both molecular (Altona Diagnostics RealStar® SARS-CoV-2 RT-PCR Kit) and antigen (FREND COVID-19 Ag) tests. Results: The area under the curve (AUC) of FREND COVID-19 Ag in all nasopharyngeal samples compared to molecular testing was 0.69 (95%CI, 0.64-0.75). At the ≥1.0 TCID50/mL manufacturer's cut-off, accuracy, sensitivity, specificity, negative (NPV) and positive (PPV) predictive values were 61.3%, 0.27, 1.00, 0.55 and 1.00, respectively. The AUC of FREND COVID-19 Ag in samples with cycle threshold (Ct) values of both SARS-CoV-2 S and E genes <29.5 was 1.00. At ≥1.0 TCID50/mL (median tissue culture infective dose per mL) manufacturer's cut-off, accuracy, sensitivity, specificity, NPV and PPV values were 99.2%, 1.00, 0.99, 1.00 and 0.95, respectively. Conclusions: FREND COVID-19 Ag could not replace routine molecular testing for achieving a definitive diagnosis of SARS-CoV-2 infection, but can be used as a surrogate test for identifying patients with higher nasopharyngeal viral load and thus greater infectious potential. © 2021 Biomedia. All rights reserved.
ABSTRACT
Background: Due to the large volume of tests needed in a relatively short time for screening and diagnosing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, antigen immunoassays may provide a potential supplement to molecular testing. This study was aimed to assess the clinical preference of DiaSorin LIAISON SARS-CoV-2 Ag chemiluminescence immunoassay. Methods: An upper respiratory specimen was collected in a series of patients referred to the Laboratory Medicine service of Pederzoli Hospital (Peschiera del Garda, Verona, Italy) for screening or diagnosis of SARS-CoV-2 infection. Nasopharyngeal samples were assayed with DiaSorin LIAISON SARS-CoV-2 Ag test and Altona Diagnostics RealStar R SARS-CoV-2 RT-PCR Kit. Results: The final study population consisted of 421 patients (median age, 48 years;227 women), 301 (71.5%) with positive result of molecular testing, and 126 (29.9%) with cycle threshold (Ct) values of both E and S genes <29.5, thus reflecting higher infectivity. The area under the curve of DiaSorin LIAISON SARS-CoV-2 Ag test 0.82 (95% CI, 0.79-0.86) for sample positivity and 0.98 for higher sample infectivity (95% CI, 0.97 to 0.99). The optimal cut-off for sample positivity was 82 TCID50/mL (0.78 sensitivity, 0.73 specificity and 77% diagnostic accuracy), whilst that for identifying samples associated with a high infective risk was 106 TCID50/mL (0.94 sensitivity, 0.96 specificity and 95% diagnostic accuracy). Conclusion: The performance of this chemiluminescence immunoassay would not permit it to replace molecular testing for diagnosing SARS-CoV-2, but may enable rapid and efficient detection of subjects with high SARS-CoV-2 viral load, who are responsible for the largest proportion of infectious clusters.