Negative PCR Research Articles

Chest CT diagnostic potential as a tool for early detection of suspected COVID-19 cases in pandemic peaks.

The emergence of severe acute respiratory syndrome coronavirus disease (COVID-19) in China at the end of 2019 caused a massive global outbreak that has become a major public health issue. Our aim was to investigate the diagnostic potential of chest CT in screening patients suspected of having COVID-19 in high-prevalence settings. This is a real-life, prospective, observational study involving 260 patients. All patients received chest CT scan at the emergency department (ED) of Kaspela University Hospital, Plovdiv, Bulgaria and RT-PCR testing for suspected COVID-19 from March 27 to December 31, 2020. COVID-19 likelihood was assessed by assigning each CT scan to a particular category of the COVID-19 Reporting and Data System (CO-RADS). IBM SPSS v. 26 was used to process the data. The male-to-female distribution ratio was 1.4:1 - 150 (57.7%) males vs. 110 (42.3%) females (p=0.014). The median age was 55 yrs (range 46-65 yrs). Discharged patients were 247 (95.0%), the rest died in the COVID-19 intensive care unit. Males were 4.13 times more likely to be diagnosed with CO-RADS≥3 score than females. Increasing age was associated with an increased likelihood of being classified with higher CO-RADS scores. The ROC curves analysis demonstrated that CO-RADS ≥3 was the optimal cutoff for discrimi-nating between a positive and negative PCR (Youden's index J=0.67), with an AUC of 0.825 (95% CI 0.72-0.93), sensitivity of 91.9% (95% CI 87.7%-95.1%), specificity of 75.0% (95% CI 53.3%-90.2%) and accuracy of 76.4% (95% CI 70.7%-81.4%). The results of this study reveal that a CT examination can provide a quick and accurate diagnosis of patients with sus-pected COVID-19 infection, whereas the PCR test is time-consuming, and the delay in receiving results can be substantial when the incidence curve begins to grow rapidly.

Steroids use in Severe/ Life Threatening COVID-19 Pneumonia: Does the Type or Dose Matter?

Objective: To compare the effect of different doses of methylprednisolone and dexamethasone on in-hospital mortality in severe COVID-19 pneumonia Methods: This retrospective chart review was done by reviewing old medical reports of patients with severe
 disease admitted to COVID-19 Intensive Care and High Dependency Unit from October 2020 to September 2021. Those with suspected COVID-19 infection (suggestive radiological findings but negative PCR for SARSCoV- 2 on at least two occasions) were excluded. Patients requiring high flow oxygen (>6 liters per minute) or higher levels of respiratory support were classified as having severe disease. We recorded the type of steroids used and the doses. Methylprednisolone in doses up to 40mg per day, or other steroids in equivalent doses, were considered low dose. Primary outcome of interest was in- hospital mortality. Results: There were 279 patients aged 52.53± 11.31 years, including 216 (77.42%) males. Mean hospital stay was 10.18± 3.13 days. During hospital stay, 96 (34.41%) patients died. Amongst patients receiving dexamethasone, 70 (44.87%) expired, whereas 26 (21.14%) out of 123 patients who received methylprednisolone expired (p

The importance of mean platelet volume lymphocyte ratio and mean platelet volume platelet ratio in COVID-19 patients

Objective: The mean platelet volume lymphocyte ratio (MPVLR) and mean platelet volume platelet ratio (MPVPR) reflect inflammation more effectively and strongly than MPV, lymphocyte, platelet, and neutrophil counts separately, we aimed to evaluate the importance of MPVLR and MPVPR in COVID-19 patients. Material and Methods: Patients with COVID-19 were divided into two groups based on whether their RT-PCR test resulted in a positive or negative result. MPVLR, MPVPR, neutrophil, lymphocyte, platelet, MPV and RDW levels were compared between these two groups. In addition, the changes in MPVLR, MPVPR, neutrophil, lymphocyte, platelet, MPV, and RDW levels before and after treatment were compared in the whole patient population. Results: There were 113 patients diagnosed with COVID-19 with a positive PCR test and 79 patients with a negative PCR test. Before treatment, the MPVLR level was 7.2±0.85 in the PCR test positive group and 5.4±0.29 in the PCR test negative group (p=0.04). Post-treatment MPVLR levels were 5.7±0.4 and 4.9±0.26 in the PCR positive and negative groups, respectively (p=0.09). The MPVPR level before treatment was 0.046±0.002 in the PCR test positive group and 0.035±0.001 in the PCR test negative group (p=0.003). After treatment, the MPVPR level was 0.040±0.002 in the PCR-positive group and 0.032±0.001 in the PCR-negative group (p=0.023) Conclusion: This study is the first to demonstrate high MPVLR and MPVPR in PCR-positive COVID-19 patients. Patients with negative PCR test and high MPVLR and MPVPR should be evaluated with radiological and clinical symptoms.

PCR for detection of Leishmania donovani from microscopically negative tissue smears of suspected patients in Gondar, Ethiopia.

As untreated visceral leishmaniasis (VL) is fatal, reliable diagnostics are pivotal for accurate treatment allocation. The current diagnostic algorithm for VL in Ethiopia, which is based on the rK39 rapid diagnostic test and microscopy of tissue smears, lacks sensitivity. This probably leads to missed cases and patients not receiving treatment. We conducted a retrospective study on stored microscopically negative spleen and bone marrow smears from suspected VL patients collected at the Leishmaniasis Research and Treatment Center (LRTC) in Gondar, northern Ethiopia between June 2019 and November 2020. Sociodemographic, clinical and treatment data were collected and samples were tested by real-time PCR targeting kinetoplast DNA. Among the 191 eligible samples (135 spleen and 56 bone marrow) with a microscopically negative and valid PCR result, 119 (62.3%) were positive by PCR, although Ct values for some were high (median 33.0). Approximately three quarters of these undiagnosed primary VL (77.3%) and relapse (69.6%) patients did not receive antileishmanial treatment. Of the 56 microscopically negative bone marrow samples, 46 (82.1%) were PCR positive, which is considerably higher compared to the microscopically negative spleen samples, for which 73 out of 135 (54.1%) were PCR positive. The odds of being PCR positive were significantly higher for bone marrow aspirates and higher when white blood cell values were lower and splenomegaly (in cm) was more pronounced. This study demonstrates that a lot of suspected VL patients remain undiagnosed and untreated. This indicates the urgent need for better diagnostics for VL in the East-African region. The outcomes of PCR positive should be closely monitored and treatment should be provided if the patient deteriorates. In resource limited settings, implementation of PCR on bone marrow aspirate smears of patients with low WBC values and splenomegaly could lead to considerable improvements in patient management.

A novel ssDNA Bidnavirus in the giant freshwater prawn Macrobrachium rosenbergii

A purported parvovirus producing circular, eosinophilic inclusions in the nuclei of tubule epithelial cells of the hepatopancreas (HP) of the giant freshwater prawn Macrobrachium rosenbergii (Mr) was first reported from Malaysia in 1990. In 2009, similar inclusions in Mr from Thailand were confirmed to contain single-stranded DNA (ssDNA). In 2021, Mr samples showing similar HP inclusions were reported to give positive PCR test results for decapod hepanhamaparvovirus (DHPV) using universal DHPV primers designed from many viral isolates of 3 penaeid shrimp species. However, the matching DNA probe revealed positive in situ hybridization (ISH) results with host HP nuclei only, and not with the intranuclear inclusions. At the time, this anomaly could not be explained. Here we describe metagenomic analysis and de novo assembly from 2016 samples of Mr with similar intranuclear inclusions that showed negative PCR test results using the DHPV universal primers. Genomes identified through metagenomics revealed a novel ssDNA genome sequence of 6723 bases that was confirmed by a single PCR amplicon. ISH using specific probes derived from this genome sequence revealed that the ssDNA was present in both grossly normal nuclei and in the intranuclear inclusions. Tests with archived DNA from the samples used in the 2021 publication yielded positive PCR results for both DHPV and the novel sequence, suggesting that the shrimp had dual infections. Phylogenetic analysis of the novel sequence revealed no relationship to any known DHPV type. Instead, the sequence closely matched the polymerase B gene in the V1 fragment from Bombyx mori bidensovirus 2 (BmBDV2) that is characterized by 2 ssDNA genome fragments and has been classified in the family Bidnaviridae. The 2 BmBDV2 fragments V1 and V2 each produce their own independent capsid protein genes and virions. In contrast, the new Mr virus is independent with a single 6723 base ssDNA genome, and we propose the name Macrobrachium hepatopancreatic bidnavirus (MHBV).

Development and validation of a nomogram to predict failure of 14-day negative nucleic acid conversion in adults with non-severe COVID-19 during the Omicron surge: a retrospective multicenter study

BackgroundWith the variability in emerging data, guidance on the isolation duration for patients with coronavirus disease 2019 (COVID-19) due to the Omicron variant is controversial. This study aimed to determine the predictors of prolonged viral RNA shedding in patients with non-severe COVID-19 and construct a nomogram to predict patients at risk of 14-day PCR conversion failure.MethodsAdult patients with non-severe COVID-19 were enrolled from three hospitals of eastern China in Spring 2022. Viral shedding time (VST) was defined as either the day of the first positive test or the day of symptom onset, whichever was earlier, to the date of the first of two consecutively negative PCR tests. Patients from one hospital (Cohort I, n = 2033) were randomly grouped into training and internal validation sets. Predictors of 14-day PCR conversion failure were identified and a nomogram was developed by multivariable logistic regression using the training dataset. Two hospitals (Cohort II, n = 1596) were used as an external validation set to measure the performance of this nomogram.ResultsOf the 2033 patients from Cohort I, the median VST was 13.0 (interquartile range: 10.0‒16.0) days; 716 (35.2%) lasted > 14 days. In the training set, increased age [per 10 years, odds ratio (OR) = 1.29, 95% confidence interval (CI): 1.15‒1.45, P < 0.001] and high Charlson comorbidity index (OR = 1.25, 95% CI: 1.08‒1.46, P = 0.004) were independent risk factors for VST > 14 days, whereas full or boosted vaccination (OR = 0.63, 95% CI: 0.42‒0.95, P = 0.028) and antiviral therapy (OR = 0.56, 95% CI: 0.31‒0.96, P = 0.040) were protective factors. These predictors were used to develop a nomogram to predict VST > 14 days, with an area under the ROC curve (AUC) of 0.73 in the training set (AUC, 0.74 in internal validation set; 0.76 in external validation set).ConclusionsOlder age, increasing comorbidities, incomplete vaccinations, and lack of antiviral therapy are risk factors for persistent infection with Omicron variant for > 14 days. A nomogram based on these predictors could be used as a prediction tool to guide treatment and isolation strategies.Graphical

COVID-19 during the Third Semester of Pregnancy: Maternal Characteristic, Possibility of Intrauterine Transmission and Neonatal Outcome in Aceh, Indonesia

Background: Pregnant women are vulnerable against COVID-19 infection due to physiological and immunological changes. COVID-19 in pregnancy affects fetal well-being with a potential for vertical infection.&#x0D; Materials and Method: This study aims to determine the incidence of vertical infection and anti-SARS-CoV-2 antibodies in infants born to mothers with positive COVID-19 infection. Amniotic fluid, swabs of the newborn's nasopharynx and oropharynx, and swabs of the placenta were examined using RT-PCR for SARS-CoV-2. Serological examination was performed by Electro-Chemiluminescence Immunoassay (ECLIA) on infant’s blood.&#x0D; Result: Four of 33 pregnant women gave birth to infants positive SARS-CoV-2 infection. RT-PCR examination of all amniotic fluid and placental swabs were negative for SARS-CoV-2. Four of 33 infants (12.1%) showed negative PCR results but positive SARS-CoV-2 antibodies, another 4 newborns (12.1%) showed positive PCR results, but no SARS-CoV-2 antibodies detected. The remaining 25 babies (75.8%) showed both negative PCR and serologic results. Conclusion: No evidence of vertical transmission found in this study.

DIAGNOSTIC ACCURACIES OF HRCT CHEST IN COMPARISON WITH RT-PCR IN DIAGNOSIS OF CORONAVIRUS DISEASE (COVID-19) IN A TERTIARY CARE HOSPITAL.

Objective:-To compare the diagnostic accuracies of HRCTchest and RT-PCR results in diagnosis of coronavirus disease (COVID-19) in a tertiary care hospital in Kolar. Methodology This was a prospective study approved by Ethical Review Committee of R L JALAPPA Hospital. From Dec 2021 to May 2022, a total of 48 CT chest were performed in this hospital. Patients aged above 18 years, irrespective of gender, who were clinically suspected of COVID-19 with at least one RT-PCR test and HRCT chest done, individuals with close contact history and admitted patients were included. Patients with interval of more than seven days between HRCT chest acquisition and rst PCR sampling, patients with pulmonary and extra pulmonary lung malignancy, chest trauma, previous chest surgery were excluded. For those patients with negative rst PCR but suspicious clinical symptoms suggesting COVID-19, a second PCR was conducted within 3 days after the rst, the result of which was taken as diagnostic gold standard. For patients with only a single RT-PCR test, the test result was taken as the diagnostic gold standard Results:- Sensitivity of HRCTin diagnosing Covid 19 was 86.49%, Specicity of HRCT in diagnosing Covid 19 was 27.27%, Positive Predictive Value of HRCT in diagnosing Covid 19 was 80.00%, Negative Predictive Value of HRCT in diagnosing Covid 19 was 37.50% and Accuracy of HRCT in diagnosing Covid 19 was 72.92%

Identification of Laboratory Biomarkers for Early Detection and Clinical Management of Post-Acute Syndrome Among Survivors of the 2013-2016 West Africa Ebola Outbreak in Sierra Leone.

The clinical management of persistent medical conditions affecting Ebola survivors, generally described as a post-Ebola syndrome, remains a public health concern. We aimed to analyze Ebola survivors' laboratory biomarkers as compared to their non-infected household relatives to identify biomarkers that could guide the identification of survivors at increased risk of developing severe at odds with the non-severe post-Ebola syndrome. Data were extracted from medical records of the Ebola survivors clinic, and we included only Ebola survivor's parameters recorded during the first baseline follow-up visit 2 weeks interval after their second negative PCR result. Moreover, household non-infected family contacts of survivors visiting the clinic during the same period were recruited as community control. The mean age of survivors was 32.65 (IQR: 15.5, 38.25) years, and Ebola IgG immunoglobulin was detected in all, thus confirming their status. The statistical significance (all p < 0.05) observed in monocyte percentage (MONO%), cluster of differentiation 4 percentage (CD4%), alanine aminotransferase (ALT), creatinine (CREA), and creatinine kinase (C-kinase) proved to be clinically significant as compared to the household relatives' group. Interestingly, the linear regression analysis indicated that the duration at ETU was negatively associated with lymphocyte percentage with a 5% lymphocyte decrease per day spent at ETU. Finally, there was a significant (p < 0.05) association between hematological (Hb, PCV, MCV, MCH), biochemical (ALT, CREA, C-kinase, T-cholesterol, triglycerides) parameters and the risk of developing severe complications. We recommend clinicians closely monitor Hb, PCV, MCV, MCH, ALT, CREA, C-kinase, T-cholesterol, triglycerides and lymphocytes as clinically relevant laboratory biomarkers to identify survivors at higher risk of developing severe post-acute syndrome upon discharge from Ebola treatment unit including headache, abdominal pain, chest pain, ocular complication, arthralgia, hearing difficulty and erectile dysfunction which can impact health-related quality of life among Ebola survivors.

Tubulointerstitial nephritis and uveitis syndrome post-COVID-19.

Tubulointerstitial nephritis and uveitis syndrome post-COVID-19.

Comparative Performance of Serological (IgM/IgG) and Molecular Testing (RT-PCR) of COVID-19 in Three Private Universities in Cameroon during the Pandemic.

COVID-19 remains a rapidly evolving and deadly pandemic worldwide. This necessitates the continuous assessment of existing diagnostic tools for a robust, up-to-date, and cost-effective pandemic response strategy. We sought to determine the infection rate (PCR-positivity) and degree of spread (IgM/IgG) of SARS-CoV-2 in three university settings in Cameroon Method: Study volunteers were recruited from November 2020 to July 2021 among COVID-19 non-vaccinated students in three Universities from two regions of Cameroon (West and Centre). Molecular testing was performed by RT-qPCR on nasopharyngeal swabs, and IgM/IgG antibodies in plasma were detected using the Abbott Panbio IgM/IgG rapid diagnostic test (RDT) at the Virology Laboratory of CREMER/IMPM/MINRESI. The molecular and serological profiles were compared, and p < 0.05 was considered statistically significant. Amongst the 291 participants enrolled (mean age 22.59 ± 10.43 years), 19.59% (57/291) were symptomatic and 80.41% (234/291) were asymptomatic. The overall COVID-19 PCR-positivity rate was 21.31% (62/291), distributed as follows: 25.25% from UdM-Bangangte, 27.27% from ISSBA-Yaounde, and 5% from IUEs/INSAM-Yaounde. Women were more affected than men (28.76% [44/153] vs. 13.04% [18/138], p < 0.0007), and had higher seropositivity rates to IgM+/IgG+ (15.69% [24/153] vs. 7.25% [10/138], p < 0.01). Participants from Bangangté, the nomadic, and the "non-contact cases" primarily presented an active infection compared to those from Yaoundé (p= 0.05, p = 0.05, and p = 0.01, respectively). Overall IgG seropositivity (IgM-/IgG+ and IgM+/IgG+) was 24.4% (71/291). A proportion of 26.92% (7/26) presenting COVID-19 IgM+/IgG- had negative PCR vs. 73.08% (19/26) with positive PCR, p < 0.0001. Furthermore, 17.65% (6/34) with COVID-19 IgM+/IgG+ had a negative PCR as compared to 82.35% with a positive PCR (28/34), p < 0.0001. Lastly, 7.22% (14/194) with IgM-/IgG- had a positive PCR. This study calls for a rapid preparedness and response strategy in higher institutes in the case of any future pathogen with pandemic or epidemic potential. The observed disparity between IgG/IgM and the viral profile supports prioritizing assays targeting the virus (nucleic acid or antigen) for diagnosis and antibody screening for sero-surveys.

Primary Aldosteronism and COVID-19-related Management, Disease Severity, and Outcomes: A Retrospective Cohort Study.

The SARS-CoV-2 virus is dependent on components of the renin-angiotensin-aldosterone system for infectivity. Primary aldosteronism (PA) is a form of secondary hypertension mediated by autonomous aldosterone production. The intersection of COVID-19 and PA, both which may involve components of the renin-angiotensin-aldosterone system, remains unknown. We assessed PA as a risk factor for COVID-19 infection and compared management, severity of disease, and outcomes during COVID-19 with a matched population of patients with essential hypertension (EH) by conducting a retrospective observational cohort study. Of the patients with PA, 81 had a negative PCR test for COVID-19, whereas 43 had a documented positive PCR test for COVID-19. Those patients with PA who tested positive for COVID-19 tended to be female (P = .08) and the majority of those with COVID-19 infection identified as non-White race (P = .02) and Hispanic ethnicity (P = .02). In a subanalysis, 24-hour urine aldosterone on initial PA diagnosis tended to be higher those in the PA group who developed COVID-19 compared with those in the PA group who did not develop COVID-19 [median (interquartile range): 36.5 (16.9, 54.3) vs 22.0 (15.8, 26.8) mcg, P = .049] and was an independent predictor of COVID-19 infection controlling for sex, race, and ethnicity. Angiotensin-converting enzyme inhibitor, angiotensin II receptor blocker, and mineralocorticoid receptor antagonist use did not differ between those patients with PA who did and did not have COVID-19 infection. Comparing those patients with PA and matched patients with EH (n = 286) who were COVID-19 PCR positive, there was a significantly higher incidence of cardiovascular complications (12 vs 2%, P = .004) in the PA vs EH group. These data begin to inform us as to whether PA should be a newly identified subpopulation at risk for COVID-19-related cardiovascular disease sequelae.

Knowledge, attitudes, behaviors, and serological status related to Chagas disease among Latin American migrants in Germany: A cross-sectional study in six German cities.

Little is known about knowledge, attitudes and behaviors concerning Chagas disease (CD) among Latin American migrants in Germany to inform public health decision making. A cross-sectional, questionnaire-based study was conducted between March 2014 and October 2019 among Latin American migrants in six cities in Germany to obtain information on migration history, socioeconomic and insurance status, knowledge about CD, potential risk factors for Trypanosoma cruzi infection, and willingness to donate blood ororgans. 168 participants completed the questionnaire. The four countries with the highest proportion of participants contributing to the study population were Colombia, Mexico, Peru and Ecuador. Before migrating to Europe, the majority of the study population resided in an urban setting in houses made of stone or concrete, had higher academic education and was integrated into the German healthcare and healthcare insurance system. The majority of all study participants were also willing to donate blood and organs and a quarter of them had donated blood previously. However, many participants lacked basic knowledge about symptoms and modes of transmission of Chagas disease. One out of 56 serologic tests (1.8%) performed was positive. The seropositive female participant born in Argentina had a negative PCR test and no signs of cardiac or other organ involvement. The study population does not reflect the population structure at risk for T. cruzi infection in endemic countries. Most participants had a low risk profile for infection with T. cruzi. Although the sample size was small and sampling was not representative of all persons at risk in Germany, the seroprevalence found was similar to studies previously conducted in Europe. As no systematic screening for T. cruzi in Latin American blood and organ donors as well as in women of child-bearing age of Latin American origin is implemented in Germany, a risk of occasional transmission of T. cruzi remains.

COVID-19 Prevention in Public Transport: A Qualitative Study

Human mobility has the potential to act as a vector for the spread of infectious agents. Efforts to prevent COVID-19 in the transportation sector must be carried out correctly. This research aims to find out the description of COVID-19 prevention behavior on public transportation. This is descriptive research with a qualitative approach. The study was conducted in Jabodetabek. The variable in this study was the prevention of COVID-19 on public transportation. This study’s informants are public transportation users selected by purposive sampling. Data were collected by conducting in-depth interviews, analyzed using the Content Analysis method, and presented in a narrative for interpretation. Atlas. ti was used to transcribe the results of in-depth interviews with informants. The findings indicate that not all modes of public transportation have the optimal level of COVID-19 prevention. Prevention of COVID-19 that is common on some public conveyance includes wearing a mask, vaccination status check, checking body temperature, hand washing facilities or hand sanitizers, and physical distancing rules. There are rules for the commuter line: do not talk while on the commuter line. Airplanes are required to have a negative PCR test. Masks and the social distancing scheme are carried out in Jaklingko and Busway (Transjakarta).COVID-19 prevention in microlets is still not optimal because keeping a distance is difficult because the reduction in passengers will affect the drivers’ income. It is hoped that the public will comply with the behavior of preventing COVID-19 in public transportation, and the government and related agencies will provide facilities and carry out monitoring and evaluation of the prevention of COVID-19 in public transportation.

SARS-CoV-2 and the ocular surface: test accuracy and viral load.

This study aimed to evaluate the pre-sence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in the ocular surface of individuals clinically suspected of coronavirus disease 2019 (COVID-19) and determine the accuracy of different approaches of molecular testing on the ocular surface based on the nasopharyngeal positivity status for COVID-19. A total of 152 individuals with suspected COVID-19 symptoms who simultaneously underwent nasopharyngeal and two different tear film collection techniques for quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR) were included. Tears were collected and randomized: one eye had the filter strip for the Schirmer test and the contralateral eye had conjunctival swab/cytology in the inferior fornix. All patients underwent slit lamp biomicroscopy. The accuracy of various ocular surface collection techniques used for the detection of SARS-CoV-2 RNA was determined. Of the 152 patients enrolled in the study, 86 (56.6%) had COVID-19 confirmed by nasopharyngeal PCR. Both tear film collection techniques detected viral particles: the Schirmer test was positive in 16.3% (14/86) and the conjunctival swab/cytology in 17.4% (15/86), with no statistically significant differences. No positive ocular tests were found among those with negative nasopharyngeal PCR tests. The overall agreement of the ocular tests was 92.7%, and in combination, the sensitivity would increase to 23.2%. The mean cycle threshold values in the nasopharyngeal, Schirmer, and conjunctival swab/cytology tests were 18.2 ± 5.3, 35.6 ± 1.4, and 36.4 ± 3.9, respectively. Compared with the nasopharyngeal test, the Schirmer (p=0.001) and conjunctival swab/cytology (p<0.001) tests had significantly different Ct values. The Schirmer (16.3%) and conjunctival swab (17.4%) tests were comparably capable of detecting SARS-CoV-2 RNA in the ocular surface by RT-PCR accurately based on nasopharyngeal status and demonstrated indistinct sensitivity and specificity. Simultaneous specimen sampling and processing from the nasopharyngeal, Schirmer, and conjunctival swab/cytology tests demonstrated significantly lower viral load in both ocular surface approaches than in the nasopharyngeal test. Ocular manifestations detected by slit lamp biomicroscopy were not associated with ocular RT-PCR positivity.

Alpha-2 macroglobulin activity in SARS-CoV-2 induced infection and in the post-COVID-19 period.

The universal proteinase inhibitor α2-macroglobulin (α₂-MG) exhibiting antiviral and immunomodulatory activities, is considered as an important participant in the infectious process. The activity of α₂-MG in the new coronavirus infection and post-covid syndrome (long COVID) has not been studied yet. We examined 85 patients diagnosed with community-acquired bilateral polysegmental pneumonia developed under conditions of a new coronavirus infection SARS-CoV-2. For assessment of the post-COVID period, 60 patients were examined 5.0±3.6 months after the coronavirus infection. Among these patients, 40 people had complications, manifested in the form of neurological, cardiological, gastroenterological, dermatological, bronchopulmonary symptoms. The control group included 30 conditionally healthy individuals with a negative PCR result for SARS-CoV-2 RNA and lack of antibodies to the SARS-CoV-2 virus. The α₂-MG activity in serum samples of patients with coronavirus infection dramatically decreased, up to 2.5% of the physiological level. This was accompanied by an increase in the activity of the α₁-proteinase inhibitor, elastase- and trypsin-like proteinases by 2.0-, 4.4- and 2.6-fold respectively as compared with these parameters in conditionally healthy individuals of the control. In the post-COVID period, despite the trend towards normalization of the activity of inhibitors, the activity of elastase-like and especially trypsin-like proteinases in serum remained elevated. In overweight individuals, the increase in the activity of trypsin-like proteinases was most pronounced and correlated with an increase in the antibody titer to the SARS-CoV-2 virus. In the post-COVID period, the α₂-MG activity not only normalized, but also exceeded the control level, especially in patients with dermatological and neurological symptoms. In patients with neurological symptoms or with dermatological symptoms, the α₂-MG activity was 1.3 times and 2.1 times higher than in asymptomatic persons. Low α₂-MG activity in the post-COVID period persisted in overweight individuals. The results obtained can be used to monitor the course of the post-COVID period and identify risk groups for complications.

Combination of granulomatosis with polyangiitis with COVID-19: difficulties in differential diagnosis and treatment

The article presents a prospective clinical observation with a fatal outcome in a patient with severe granulomatosis with polyangiitis (GPA) complicated by COVID 19 on the background of immunosuppression. At the onset of the disease, there were difficulties in making a diagnosis; differential diagnosis between eosinophilic granulomatosis with polyangiitis (EGPA) and GPA was carried out. Against the background of combined therapy — the use of high and ultra-high doses of glucocorticoids (GC) and cytostatic drugs, a short-term improvement was noted. However, within a year and a half of treatment, it was not possible to achieve remission of the disease. Taking into account the ineffectiveness of the therapy, by the decision of the medical commission, the patient was prescribed genetically engineered biological therapy rituximab (RTM). There was a decrease in the activity of the disease, the achievement of depletion of B-cells. During the pandemic period, despite observing the isolation regime, the patient fell ill with a coronavirus infection. Immunosuppression contributed to severe infection. After the infection was treated, the activity of vasculitis increased, which required the repeated administration of RTM for health reasons. During the therapy it was short-term stabilization of the condition. But after short period — fever, an increase in respiratory failure, the development of neutropenia. Taking into account the initial lesion of the lungs in the patient, differential diagnostics was carried out between the complications of coronavirus infection and the activity of the underlying disease. Despite anti-inflammatory, anticoagulant, antibacterial therapy, the patient's condition progressively worsened, respiratory failure increased, hemoptysis appeared. Spontaneous pneumothorax on the right was diagnosed, pneumomediastinum. After repeated negative PCR results, the causative agent of SARS COV-2 was again detected in smears. The immediate cause of death, according to clinical observation and autopsy, was severe respiratory failure, thrombosis in small vessels of the lungs in a patient with a new coronavirus infection against the background of immunosuppression for GPA.

A Laboratory-Based Study on Multiple Biomarker Testing in the Diagnosis of COVID-19-Associated Pulmonary Aspergillosis (CAPA): Real-Life Data

(1) Background: Coronavirus disease 2019 (COVID-19)-associated pulmonary aspergillosis (CAPA) raises concerns to contribute to an increased mortality. The incidence of CAPA varies widely within hospitals and countries, partly because of difficulties in obtaining a reliable diagnosis. (2) Methods: Here, we assessed Aspergillus culture-positive and culture-negative respiratory tract specimens via direct fungal microscopy (gold standard) and compared the results with galactomannan enzyme immunoassay (GM-EIA) and Aspergillus PCR. (3) Results: 241 respiratory samples from patients suffering from SARS-CoV-2 pneumonia were evaluated. Results showed both diagnostic tools, Aspergillus PCR and GM-EIA, to be positive or negative displaying a sensitivity of 0.90, a specificity of 0.77, a negative predictive value (NPV) of 0.95, and a positive predictive value (PPV) of 0.58 in Aspergillus sp. culture and microscopic-positive specimens. Non-bronchoalveolar lavage (BAL) samples, obtained within a few days from the same patient, showed a high frequency of intermittent positive or negative GM-EIA or Aspergillus PCR results. Positivity of a single biomarker is insufficient for a proper diagnosis. A broad spectrum of Aspergillus species was detected. (4) Conclusions: Our study highlights the challenges of combined biomarker testing as part of diagnosing CAPA. From the results presented, we highly recommend the additional performance of direct microscopy in respiratory specimens to avoid overestimation of fungal infections by applying biomarkers.

Therapeutic effect and safety of ozone therapy in sars cov-2 patients. Cuban experience

SARS-Cov2, the virus causing COVID-19, is distributed globally since December 2019, classified as a pandemic by the World Health Organization. The virus is known to cause Acute Respiratory Distress Syndrome (ARDS). Cytokine storm is considered to be one of the major causes of ARDS and multiple-organ failure. Due to the high lethality of SARS-CoV2 infections and its economic and social impact, it is necessary to seek new therapeutic procedures. It has been demonstrated that ozone therapy produces a significant improvement in blood flow and oxygenation of ischemic tissues. Also, ozone can achieve an equilibrium between Nrf2 and NF-?B factors, modulating oxidative stress and the expression of pro-inflammatory cytokines. Levels of reduced glutathione (GSH), the key antioxidant guardian in all tissues, could be critical in extinguishing the exacerbated inflammation that triggers organ failure in COVID-19. The reduced levels of GSH are clinically associated with severe disease. It is well documented that ozone activates the level of GSH. In clinical studies, ozone has a significant role in the treatment of pulmonary and vascular diseases. An exploratory, controlled, open and monocentric study was carried out in 32 patients with mild to moderate symptoms, distributed at random in two groups of 16 patients each. The first group received rectal ozone therapy (ozone concentration between 25 and 40 mg/L and volume between 100 and 200 ml, twice per day) together with conventional treatment with Kaletra Lopinavir/Ritonavir), chloroquine and/or Heberferon®, and the second group only conventional treatment. The result of the RT-PCR and the evolution of the clinical signs were the primary efficacy variables. Secondary efficacy variables were ferritin, C-reactive protein, oxidative stress biomarkers, cellular indicators of inflammation, biochemical and hematological variables, and the presence of adverse effects. Patients (81%) who received rectal ozone therapy had a negative RT-PCR after the tenth ozone therapy application (after 5 days), with significant differences from the control group at the same time. In all patients, improvements in the levels of C-reactive protein and cellular indicators of inflammation were achieved at the end of the treatment. Ozone treatment significantly increases GSH levels compared to the control group, but not other REDOX markers. The treatments were safe and welltolerated.&#x0D; Also, a pilot, open-label, phase IV clinical trial study was performed in 40 SARS-CoV-2 convalescent patients (with symptoms extending beyond three weeks since negative PCR test) distributed at random into 2 groups. One group was treated with BIOPLA (dietary complement based on a concentrated protein - mineral elaborated from human placental after born, 1 capsule of 400 mg during 30 days) and the ozone group treated with rectal ozone application (ozone concentration between 20 and 35 mg/L and volume between 150 and 200 ml, 20 sessions, one per day) + BIOPLA. Both groups were treated and followed for one month. Twenty individuals supposedly healthy were enrolled and evaluated as the control group.&#x0D; Effect evaluation showed beneficial improvement in both treated groups resulting in 85% of patients for BIOPLA plus RIO3T and 37% for BIOPLA (p

Time to negative PCR conversion amongst high-risk patients with mild-to-moderate Omicron BA.1 and BA.2 COVID-19 treated with sotrovimab or nirmatrelvir

ObjectivesOur aim was to compare the clinical and virological outcomes in Omicron BA.1- and BA.2-infected patients who received sotrovimab with those in patients who received nirmatrelvir for the prevention of severe COVID-19. MethodsIn this multi-centric, prospective ANRS 0003S CoCoPrev cohort study, patients at a high risk of progression of mild-to-moderate BA.1 or BA.2 COVID-19 who received sotrovimab or nirmatrelvir were included. The proportion of patients with progression to severe COVID-19, time between the start of treatment to negative PCR conversion, SARS-CoV-2 viral decay, and characterization of resistance variants were determined. A multi-variable Cox proportional hazard model was used to determine the time to negative PCR conversion and a mixed-effect model for the dynamics of viral decay. ResultsAmongst 255 included patients, 199 (80%) received ≥3 vaccine doses, 195 (76%) received sotrovimab, and 60 (24%) received nirmatrelvir. On day 28, new COVID-19-related hospitalization occurred in 4 of 193 (2%; 95% CI, 1–5%) sotrovimab-treated patients and 0 of 55 nirmatrelvir-treated patients (p 0.24). One out of the 55 nirmatrelvir-treated patients died (2%; 95% CI, 0–10%). The median time to negative PCR conversion was 11.5 days (95% CI, 10.5–13) in the sotrovimab-treated patients vs. 4 days (95% CI, 4–9) in the nirmatrelvir-treated patients (p < 0.001). Viral decay was faster in the patients who received nirmatrelvir (p < 0.001). In the multi-variable analysis, nirmatrelvir and nasopharyngeal PCR cycle threshold values were independently associated with faster conversion to negative PCR (hazard ratio, 2.35; 95% CI, 1.56–3.56; p < 0.0001 and hazard ratio, 1.05; 95% CI, 1.01–1.08; p 0.01, respectively). ConclusionsEarly administration of nirmatrelvir in high-risk patients compared with that of sotrovimab was associated with faster viral clearance. This may participate to decrease transmission and prevent viral resistance.