The aim of our study was to evaluate the diagnostic performance of two antigen rapid diagnostic tests (Ag‐RDTs) to diagnose severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection. We ...evaluated Panbio and SD‐Biosensor Ag‐RDTs. We employed 186 polymerase chain reaction (PCR) negative samples to evaluate the specificity and 170 PCR positive samples to assess the sensitivity. We evaluated their sensitivity according to Cycle threshold (C
t) values and days post onset of symptoms (d.p.o.). Tests were compared using the McNemar's test. Agreement was evaluated using the kappa score. Specificity was 100% for Panbio and 97.3% for SD‐Biosensor. Sensitivity for samples with C
t ≤ 20 was 100% for both assays and for samples with C
t = 20–25 was 93.0% (Panbio) and 95.3% (SD‐Biosensor) (p = 1.000). Sensitivity decreased for samples wit C
t = 25–30 (Panbio: 41.3%, SD‐Biosensor: 52.2%, p = 0.125) and samples with C
t ≥ 30 (Panbio: 5.0%, SD‐Biosensor: 17.5%, p = 0.063). Sensitivity within seven d.p.o. was 87.7% for Panbio and 90.4% for SD‐Biosensor and notably decreased after seven d.p.o. Agreement with PCR was excellent for high viral load samples (C
t ≤ 25): Panbio, 98.9%, kappa = 0.974; SD‐Biosensor, 97.4%, kappa = 0.940. Agreement between Ag‐RDTs was excellent (94.9%, kappa = 0.882). Panbio and SD‐Biosensor Ag‐RDTs showed excellent agreement and diagnostic performance results for samples with high viral loads (C
t ≤ 25) or samples within seven d.p.o.
Highlights
Panbio and SD‐Biosensor Ag‐RDTs are reliable to diagnose SARS‐CoV‐2 infection.
They showed high specificity: 100% (Panbio) and 97.3% (SD‐Biosensor).
Sensitivity for samples with Ct ≤ 20 was 100% and for samples with Ct ≤ 25 was over 93%.
Their sensitivity was over 87% within 7 days after symptoms onset. Agreement between them was excellent (agreement = 94.9%, kappa = 0.882).
Oocyte activation via dual inhibition of protein synthesis and phosphorylation has improved in vitro embryo production in different mammalian species. In this study, we evaluated the effects of the ...combination of cycloheximide (CHX), dimethyl amino purine (DMAP), and anisomycin (ANY) on the activation of bovine oocytes, particularly on dynamics of MPF and MAPKs, embryonic developmental potential, and quality. The results showed that the cleavage and blastocyst rates, as well as levels of CCNB1, CDK1, p-CDK1Thr161, and p-CDK1Thr14-Tyr15, were similar among groups; ANY and ANY + CHX reduced the expression of ERK1/2 compared to DMAP-combinations (p < 0.05), whereas ANY + DMAP, CHX + DMAP, and ANY + CHX + DMAP reduced p-ERK1/2 compared to ANY and ANY + CHX treatments (p < 0.05). The quality of blastocysts in terms of cell counts, their allocation, and the numbers of TUNEL-positive cells did not differ among groups. However, transcript levels of POU5F1 were higher in embryos derived from ANY + CHX + DMAP treatment compared to other groups, while expression levels of CDX2 did not show differences. In addition, the BCL2A1/BAX ratio of the ANY + CHX + DMAP treatment was significantly low compared to the ANY treatment (p < 0.05) and did not differ significantly from the other treatments. In conclusion, oocyte activation by dual inhibition of protein synthesis and phosphorylation induces MPF inactivation without degradation of CCNB1, while MAPK inactivation occurs differentially between these inhibitors. Thus, although the combined use of these inhibitors does not affect early developmental competence in vitro, it positively impacts the expression of transcripts associated with embryonic quality.
•The new antigen test is an indispensable tool in the control of the pandemic due to its adequate sensitivity and specificity.•The implementation of the point of care technique in primary care is ...feasible and has good results.•Results of the antigen techniques are determined by an onset of symptoms inferior to five days and a CT below 27 in PCR.
RT-qPCR is the current recommended laboratory method to diagnose SARS-CoV-2 acute infection, several factors such as requirement of special equipment, time consuming, high cost and skilled staff limit the use of these techniques. A more rapid and high-throughput method is essential.
We analyzed clinical data and nasopharyngeal samples, collected during September 2020, from patients attended at the emergency department of a secondary hospital and in two primary healthcare centers in Madrid. The performance of the Panbio™ COVID-19 AG Rapid Test Device for the detection of SARS-CoV-2 antigen was compared to RT-qPCR.
255 nasopharyngeal swabs, including 150 from the emergency department and 105 from primary helthcare centers, were tested. 184 patients were symptomatic (72.1 %). Amongst the 60 positive RT-qPCR samples, 40 were detected by the rapid antigen test, given an overall sensitivity of 73.3 %. All the samples detected positive with the rapid antigen test were also positive with RT-qPCR. The median cycle threshold was 23.28 (IQR 18.5–30.16). Patients with less than seven days onset of symptoms showed a higher viral load, and sensitivity for rapid antigen test (86.5 %), compared to those with more days (sensitivity of 53.8 %)(p < 0.004).
The rapid antigen test evaluated in this study showed a high sensitivity and specificity in samples obtained during the first week of symptoms and with high viral loads. This assay seems to be an effective strategy for controlling the COVID-19 pandemic for the rapid identification and isolation of SARS-CoV-2 infected patients.
The objectives of our study were to describe the characteristics of patients with
candidemia and to perform an in-depth microbiological characterization of isolates and compare them with those of ...patients with
candidemia. We described the risk factors and outcomes of 22 patients with candidemia caused by the
complex. Incident isolates were identified using molecular techniques, and susceptibility to fluconazole, anidulafungin, and micafungin was studied. Biofilm formation was measured using the crystal violet assay (biomass production) and the XTT reduction assay (metabolic activity), and virulence was studied using the
model. Biofilm formation was compared with that observed for
The main conditions predisposing to infection were malignancy (68%), immunosuppressive therapy (59%), and neutropenia (18%). Clinical presentation of candidemia was less severe in patients infected by the
complex than in patients infected by
, and 30-day mortality was lower in
patients (13.6% versus 33.9%, respectively;
= 0.049). Isolates were identified as
(
= 17) and
(
= 5). The isolates produced biofilms with low metabolic activity and moderate biomass. The
model showed that
was less virulent than
(mean of 6 days versus 1 day of survival, respectively;
< 0.001). Patients with candidemia caused by the
complex had severe and debilitating underlying conditions. Overall, the isolates showed diminished susceptibility to fluconazole and echinocandins, although poor biofilm formation and the low virulence were associated with a favorable outcome.
•SARS-CoV-2 induces the expression of innate immune genes in the nasopharynx.•Expression of immune genes in the nasopharynx was associated with COVID-19 severity.•High plasminogen activator urokinase ...receptor and low chemokine C-X-C motif ligand 10 expression levels were related to COVID-19 pneumonia.•Low expression of interferon-stimulated gene 15, retinoic acid-inducible gene I, C-C motif ligand 5, and chemokine C-X-C motif ligand 10 was related to severe pneumonia.
We analyzed the expression of inflammatory and antiviral genes in the nasopharynx of SARS-CoV-2 infected patients and their association with the severity of COVID-19 pneumonia.
We conducted a cross-sectional study on 223 SARS-CoV-2 infected patients. Clinical data were collected from medical records, and nasopharyngeal samples were collected in the first 24 hours after admission to the emergency room. The gene expression of eight proinflammatory/antiviral genes (plasminogen activator urokinase receptor PLAUR, interleukin IL-6, IL-8, interferon IFN-β, IFN-stimulated gene 15 ISG15, retinoic acid-inducible gene I RIG-I, C-C motif ligand 5 CCL5, and chemokine C-X-C motif ligand 10 CXCL10) were quantified by real-time polymerase chain reaction. Outcome variables were: (i) pneumonia; (ii) severe pneumonia or acute respiratory distress syndrome. Statistical analysis was performed using multivariate logistic regression analyses.
We enrolled 84 mild, 88 moderate, and 51 severe/critical cases. High expression of PLAUR (adjusted odds ratio aOR = 1.25; P = 0.032, risk factor) and low expression of CXCL10 (aOR = 0.89; P = 0.048, protective factor) were associated with pneumonia. Furthermore, lower values of ISG15 (aOR = 0.88, P = 0.021), RIG-I (aOR = 0.87, P = 0.034), CCL5 (aOR = 0.73, P <0.001), and CXCL10 (aOR = 0.84, P = 0.002) were risk factors for severe pneumonia/acute respiratory distress syndrome.
An unbalanced early innate immune response to SARS-CoV-2 in the nasopharynx, characterized by high expression of PLAUR and low expression of antiviral genes (ISG15 and RIG-I), and chemokines (CCL5 and CXCL10), was associated with COVID-19 severity.
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Background
Higher expression of olfactomedin‐4 (OLFM4), a gene regulated by nuclear factor‐kappa B (NF‐κB), has been related to a higher risk of organ failure and death in patients with septic shock. ...We aimed to evaluate the association between OLFM4 single nucleotide polymorphisms (SNPs) and septic shock‐related death in 175 patients who underwent major surgery, as well as its performance in predicting mortality.
Materials and methods
We carried out a retrospective study. A total of seven OLFM4 SNPs were genotyped by Agena Bioscience's MassARRAY platform. Statistical analysis was performed by Kaplan‐Meier and Cox regression tests. The diagnostic performance for predicting septic shock‐related death was evaluated by the area under the receiver‐operating characteristic (AUROC) curve.
Results
Patients with rs17552047 A allele and rs1891944 TT genotype had higher survival than patients with rs17552047 G allele (P‐value = .024) and patients with rs1891944 CC/CT genotype (P‐value = .038). However, only rs17552047 was associated with a lower risk of death under an additive inheritance model (adjusted hazard ratio aHR = 0.44, 95% CI = 0.27‐0.71). The multivariate model with the most significant clinical variables (lactate, chronic kidney disease, peritonitis, heart disease and elective surgery) showed an AUROC of 0.776 for predicting septic shock‐related death. When we added the OLFM4 rs17552047 SNP to the previous model, the AUROC was 0.811 and was close to reaching significant differences with the previous model (P‐value = .065).
Conclusion
OLFM4 rs17552047 A allele predicts septic shock survival in patients who underwent major surgery. Furthermore, rs17552047, together with clinical variables, could be useful to predict the outcome of septic shock.
We carried out a retrospective exploratory study on 173 patients who underwent major surgery and developed septic shock after surgery. Our findings suggest that CEACAM7 rs1001578, rs10409040, and ...rs889365 polymorphisms could influence septic shock-related death in individuals who underwent major surgery.
A limited number of longitudinal studies have examined the symptoms associated with long-COVID-19. We conducted an assessment of symptom onset, severity and patient recovery, and determined the ...percentage of patients who experienced reinfection up to 2 years after the initial onset of the disease. Our cohort comprises 377 patients (>=18 years) with laboratory-confirmed COVID-19 in a secondary hospital (Madrid, Spain), throughout March 3-16, 2020. Disease outcomes and clinical data were followed-up until August 12, 2022. We reviewed the evolution of the 253 patients who had survived as of April 2020 (67.1%). Nine died between April 2020 and August 2022. A multivariate regression analysis performed to detect the risk factors associated with long-COVID-19 revealed that the increased likelihood was associated with chronic obstructive lung disease (OR 14.35, 95% CI 1.89-109.09; 'p' = 0.010), dyspnea (5.02, 1.02-24.75; 'p' = 0.048), higher LDH (3.23, 1.34-7.52; 'p' = 0.006), and lower D-dimer levels (0.164, 0.04-0.678; 'p' = 0.012). Reinfected patients ('n' = 45) (47.8 years; 39.7-67.2) were younger than non-reinfected patients (64.1 years; 48.6-74.4)) ('p' < 0.001). Patients who received a combination of vaccines exhibited fewer symptoms (44.4%) compared to those who received a single type of vaccine (77.8%) ('p' = 0.048). Long-COVID-19 was detected in 27.05% (66/244) of patients. The early detection of risk factors helps predict the clinical course of patients with COVID-19. Middle-aged adults could be susceptible to reinfection, highlighting the importance of prevention and control measures regardless of vaccination status.
COVID-19 can course with respiratory and extrapulmonary disease. SARS-CoV-2 RNA is detected in respiratory samples but also in blood, stool and urine. Severe COVID-19 is characterized by a ...dysregulated host response to this virus. We studied whether viral RNAemia or viral RNA load in plasma is associated with severe COVID-19 and also to this dysregulated response.
A total of 250 patients with COVID-19 were recruited (50 outpatients, 100 hospitalized ward patients and 100 critically ill). Viral RNA detection and quantification in plasma was performed using droplet digital PCR, targeting the N1 and N2 regions of the SARS-CoV-2 nucleoprotein gene. The association between SARS-CoV-2 RNAemia and viral RNA load in plasma with severity was evaluated by multivariate logistic regression. Correlations between viral RNA load and biomarkers evidencing dysregulation of host response were evaluated by calculating the Spearman correlation coefficients.
The frequency of viral RNAemia was higher in the critically ill patients (78%) compared to ward patients (27%) and outpatients (2%) (p < 0.001). Critical patients had higher viral RNA loads in plasma than non-critically ill patients, with non-survivors showing the highest values. When outpatients and ward patients were compared, viral RNAemia did not show significant associations in the multivariate analysis. In contrast, when ward patients were compared with ICU patients, both viral RNAemia and viral RNA load in plasma were associated with critical illness (OR CI 95%, p): RNAemia (3.92 1.183-12.968, 0.025), viral RNA load (N1) (1.962 1.244-3.096, 0.004); viral RNA load (N2) (2.229 1.382-3.595, 0.001). Viral RNA load in plasma correlated with higher levels of chemokines (CXCL10, CCL2), biomarkers indicative of a systemic inflammatory response (IL-6, CRP, ferritin), activation of NK cells (IL-15), endothelial dysfunction (VCAM-1, angiopoietin-2, ICAM-1), coagulation activation (D-Dimer and INR), tissue damage (LDH, GPT), neutrophil response (neutrophils counts, myeloperoxidase, GM-CSF) and immunodepression (PD-L1, IL-10, lymphopenia and monocytopenia).
SARS-CoV-2 RNAemia and viral RNA load in plasma are associated with critical illness in COVID-19. Viral RNA load in plasma correlates with key signatures of dysregulated host responses, suggesting a major role of uncontrolled viral replication in the pathogenesis of this disease.
•Alltest lateral flow immunoassay is reliable to diagnose SARS-CoV-2 infection.•This immunoassay showed a specificity of 100 % and a sensitivity of 64 %.•The sensitivity increased up to 88 % from 14 ...days after symptoms onset.•Alltest is useful in patients with suspected SARS-CoV-2 pneumonia but negative PCR.•The test was positive in 89 % of patients with pneumonia and negative PCR.
SARS-CoV-2 infection diagnosis is challenging in patients from 2 to 3 weeks after the onset of symptoms, due to the low positivity rate of the PCR. Serologic tests could be complementary to PCR in these situations. The aim of our study was to analyze the diagnostic performance of one serologic rapid test in COVID-19 patients.
We evaluated a lateral flow immunoassay (AllTest COVID-19 IgG/IgM) which detects IgG and IgM antibodies. We validated the serologic test using serum samples from 100 negative patients (group 1) and 90 patients with COVID-19 confirmed by PCR (group 2). Then, we prospectively evaluated the test in 61 patients with clinical diagnosis of pneumonia of unknown etiology that were negative for SARS-CoV-2 by PCR (group 3).
All 100 patients from group 1 were negative for the serologic test (specificity = 100 %). Regarding group 2 (PCR-positive), the median time from their symptom onset until testing was 17 days. For these 90 group-2 patients, the test was positive for either IgM or IgG in 58 (overall sensitivity = 64.4 %), and in patients tested 14 days or more after the onset of symptoms, the sensitivity was 88.0 %. Regarding the 61 group-3 patients, median time after symptom onset was also 17 days, and the test was positive in 54 (88.5 % positivity).
Our study shows that Alltest lateral flow immunoassay is reliable as a complement of PCR to diagnose SARS-CoV-2 infection after 14 days from the onset of symptoms and in patients with pneumonia and negative PCR for SARS-CoV-2.
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