Viral RNA load in plasma is associated with critical illness and a dysregulated host response in COVID-19.

BACKGROUND: 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. METHODS: 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. RESULTS: 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). CONCLUSIONS: 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.

Use of Saliva Swab for Detection of Influenza Virus in Patients Admitted to an Emergency Department.

Nasopharyngeal (NP) specimens are commonly used for the detection of influenza, but saliva swabs are easier to obtain and cause less discomfort to the patients. The objective of this study was to evaluate the usefulness of saliva swab specimens for the diagnosis of influenza compared with NP specimens. Influenza virus detection rate in saliva and NP swabs was compared in adult patients admitted to an emergency department from January to March 2020, using the Xpert Xpress Flu/respiratory syncytial virus (RSV) test. Cycle threshold (CT) values were evaluated in all the cases. Among the 82 patients recruited, 19 had an influenza-positive diagnostic test result (11 influenza A and 8 influenza B). Overall, the agreement between saliva and NP swabs results was 97.6% (80/82; κ = 0.929; 95% confidence interval [CI], 0.832 to 1.0). There was no significant difference in the influenza detection rate between saliva swab and NP specimens (20.7% [17/82] versus 23.2% [19/82]; P = 0.5). There were only two discordant results (influenza B in an NP and false negative in a saliva sample). Manual inspection of the amplification curves showed that influenza RNA had been amplified in saliva with high CTs (CT of 40) that the test reported as a negative result. The overall sensitivity and specificity for saliva was 89.5% (73.0% to 100%) and 100% (99.2% to 100%), respectively. In all the cases, the same influenza virus (A/B) was detected. Median CT values were significantly lower in NP (31; interquartile range [IQR], 21.0 to 32.0) than in saliva (33; IQR, 23.0 to 38.0) (P = 0.001) specimens. Saliva swabs have high sensitivity and specificity for the detection of influenza virus by the Xpert Xpress Flu/RSV test and a high overall agreement and CT correlation with NP specimens. Saliva swab is a feasible specimen type for influenza testing that might be easily self-collected with minimal equipment and discomfort. IMPORTANCE Early detection of influenza virus is important for guiding antiviral and antibacterial treatment for infection control and public health measures. We have observed that saliva swab specimens have high sensitivity and specificity for the detection of influenza by the Xpert Xpress Flu/respiratory syncytial virus (RSV) test and high overall agreement and CT correlation with nasopharyngeal specimens. Saliva swab may therefore be a feasible specimen type for influenza testing that can be easily self-collected with minimal equipment and discomfort.