Nasal Specimens Research Articles

EPH66 Epidemiological Population Study of Sars-Cov-2 in Lake County, Illinois: Contact: Health-Related Quality of Life Assessment

The prospective, longitudinal, community-based CONTACT study aimed to improve our understanding of COVID-19 immunity, and other characteristics related to SARS-CoV-2 long-term, including the assessment of health-related quality of life (HRQoL) at baseline and over time by infection status. Participants living or working in Lake County, IL were recruited between November 2020 and January 2021. At baseline and follow up visits (3-, 6-, and 9-Months-M-), participants self-reported their occupational exposure, COVID-19 vaccination status and provided nasal and blood serum specimens for molecular (RT-PCR) and serologic (IgG) testing to detect current or previous SARS-CoV-2 infection. HRQoL questionnaires EQ-5D-5L were completed online approximately within two weeks post-testing (at 0.5, 3.5, 6.5, and 9.5 months) after results were communicated. EQ-5D-5L information was described and stratified by COVID-19 status at baseline, 3M, 6M and 9M – software: SAS®-v9.4. Data from 1008 participants were analyzed. Participants testing positive to COVID-19 were 56/952, 48/751, 40/693, and 19/654, respectively, at baseline, 3M, 6M, and 9M. Of the five domains of EQ-5D-5L, a higher percentage of participants who tested positive for COVID-19 reported having no anxiety or depression versus those who tested negative: at baseline (55.4% [31/56] vs 50.5% [481/952]); 3M (68.8% [33/48] vs. 56.3% [423/751]); 6M (67.5% [27/40] vs. 56.3% [390/693]); and 9M (73.7% [14/19] vs. 60.4% [395/654]). Median Visual Analogue Scale (VAS) score was at least 2 points higher at all time- points for participants who tested positive except at last visit (baseline: 89.0 vs. 87.0; 3M: 88.0 vs. 86.0; 6M: 87.5 vs. 85.0; 9M: 85.0 vs. 87.0) This analysis provides insight into participant HRQoL burden at enrollment and over time when a positive test to COVID-19 was communicated. At all time-points, anxiety or depression was experienced by more participants who tested negative versus those who didn´t.

The association between allergic rhinitis and airway dysfunction and nasal endothelial damage and oxidative stress.

Although lower airway hyperresponsiveness is present in approximately one in three patients with allergic rhinitis (AR), the underlying mechanism remains unclear. To evaluate nasal patency and pulmonary functions in AR independently of the presence of asthma and to investigate the relationships between these and nasal oxidative stress parameters and endothelial damage. Seventy adolescents with AR (AR group - 27 with asthma and 43 without asthma) and 30 healthy controls (HC group) were included in this prospective, cross-sectional study. Endocan and oxidative biomarkers [total oxidant status (TOS), total antioxidant status (TAS), and oxidative stress index (OSI)] in nasal lavage fluid specimens; peak nasal inspiratory flow (PNIF); fractional exhaled nitric oxide (FeNO), and impulse oscillometry (zR5, zR20, and R5-20 for resistance and zX5 and zX20 for reactance) were investigated. Nasal endocan, TOS, and OSI values were higher in the AR group and TAS in the HC group. There was no difference between AR groups with and without asthma in terms of nasal endocan and oxidative biomarkers. FeNO levels and airway resistance (zR5, zR20, and R5-20) were higher in the AR group than in the HC group. However, there was no difference between the groups in PNIF. X5 was higher among the AR without asthma than in the other groups. Correlation between OSI and R5-20 was observed in the AR group. In the linear regression model, (logged) OSI was significantly predicted (logged) R5-20. The airways of adolescents with AR without asthma were as much affected as those of the AR with asthma, and this effect was associated with nasal endothelial damage and an increase in oxidative stress.

Meningoencephalitis Secondary To Cerebrospinal Fluid Leakage From Nasopharyngeal COVID-19 Swab (P6-10.006)

<h3>Objective:</h3> NA <h3>Background:</h3> The WHO regarded the novel coronavirus as an outbreak on January 2020. The preferable initial testing to detect SARS-CoV-2 RNA was from the upper respiratory tract using nucleic acid amplification testing (NAAT) through nasopharyngeal, oropharyngeal, nasal mid-turbinate, or anterior nasal specimen. Although specimen collection is considered safe and adverse events are rare, some reported cases of cerebrospinal fluid leakage were attributed to pharyngeal swab procedures. Therefore, increased awareness of nasal and pharyngeal swab complications is necessary. <h3>Design/Methods:</h3> A 36 years old female known to have chronic idiopathic intracranial hypertension brought to the emergency department with a history of fever, altered level of consciousness, and generalized tonic-clonic seizure. She had unilateral rhinorrhea for three months following a nasopharyngeal COVID-19 swab as part of an in-vitro fertilization workup. Upon assessment in the ED, she was stuporous due to sedation effect. There was no clear rhinorrhea, no papilledema on the fundus exam, and the remaining cranial nerves examination was unremarkable. She had positive meningeal signs. A cloudy cerebrospinal fluid analysis revealed findings of bacterial meningitis with high WBC and protein. She was started on empirical antimeningeal coverage. MRI brain showed a small CSF containing space at the left inferior frontal in the olfactory groove, which is continuous with the adjacent sulci concerning for the CSF leak site. Beta-transferrin was requested by the otolaryngology team and came back positive. After fourteen days of antibiotics, she regained her baseline cognitive and functional status then was discharged in stable condition with close follow-up with neurology, otolaryngology, and MRI cisternography. <h3>Results:</h3> NA <h3>Conclusions:</h3> Our case emphasizes that awareness of potential adverse events related to nasopharyngeal swabs that became part of daily clinical practice and hospital policies must be improved. It also supports existing scattered evidence that an existing idiopathic intracranial hypertension can be associated with cerebrospinal fluid leakage. <b>Disclosure:</b> Dr. Alnafisah has nothing to disclose. Dr. Altowairqi has nothing to disclose.

Maternal transfer of IgA and IgG SARS-CoV-2 specific antibodies transplacentally and via breast milk feeding.

Although there have been many studies on antibody responses to SARS-CoV-2 in breast milk, very few have looked at the fate of these in the infant, and whether they are delivered to immunologically relevant sites in infants. Mother/infant pairs (mothers who breast milk fed and who were SARS-CoV-2 vaccinated before or after delivery) were recruited for this cross-sectional study. Mother blood, mother breast milk, infant blood, infant nasal specimen, and infant stool was tested for IgA and IgG antibodies against SARS-CoV-2 spike trimer. Thirty-one mother/infant pairs were recruited. Breast milk fed infants acquired systemic anti-spike IgG antibodies only if their mothers were vaccinated antepartum (100% Antepartum; 0% Postpartum; P<0.0001). Breast milk fed infants acquired mucosal anti-spike IgG antibodies (in the nose) only if their mothers were vaccinated antepartum (89% Antepartum; 0% Postpartum; P<0.0001). None of the infants in either group had anti-spike IgA in the blood. Surprisingly, 33% of the infants whose mothers were vaccinated antepartum had high titer anti-spike IgA in the nose (33% Antepartum; 0% Postpartum; P = 0.03). Half-life of maternally transferred plasma IgG antibodies in the Antepartum infant cohort was ~70 days. Vaccination antepartum followed by breast milk feeding appears to be the best way to provide systemic and local anti-SARS-CoV-2 antibodies for infants. The presence of high titer SARS-CoV-2-specific IgA in the nose of infants points to the potential importance of breast milk feeding early in life for maternal transfer of mucosal IgA antibodies. Expectant mothers should consider becoming vaccinated antepartum and consider breast milk feeding for optimal transfer of systemic and mucosal antibodies to their infants.

Определение сроков декальцинации образцов грудины, коленного сустава и препарата носовых ходов мышей, песчанок и хомячков

Procedure of decalcification necessary for histology of bone tissue. Decalcification is the procedure of removing minerals from the bone organic matrix. Solutions of varying acids usually used as a decalcification solution. Whole bone or piece that had been cut out preliminary are can be sampled for decalcification. Study of a knee joint and nasal cavity specimens usually uses in preclinical research. Also, for the study of bone marrow, a sternum often is sampled. The cutting out of small piece often leads to the destruction of the sample, because of their small size. Thus, whole maxilla, whole sternum and knee joint to include femur, fibula and tibia are more often sampled for research. This article describe a time of decalcification above-mentioned specimen that were sampled from mice, gerbils and hamsters. Solutions of formic and hydrochloric acids were used as the decalcification solution. The decalcification solution were changed every other day. The severity of decalcifications and specimen readiness for histology procedure were evaluated by gravimetric method. We found that the trends in mass change and decalcification time change were the same for all animal types noted above. In none case, the absolute severity of decalcification was reached, within 10 days. However, the severity of decalcification of whole specimens of bone was sufficient to make a quality histological slide. All specimens achieved the sufficient severity of decalcification on 4th day. This period allows to evaluate the morphology of the respiratory and olfactory epithelium of the nasal cavity, joint cartilage and bone marrow cells. The period of decalcification can be extended to 7 days, if anatomic and topographic features and them the best quality more important. The period of decalcification more 7 days leaded to irreversible change of cell morphology. Cells had poor detalization, cell nuclei were stained with hematoxiline badly, cell borders were unclear.

Nasal Sesamoid Cartilages: Fact or Fiction? An Anatomical Study of the Lateral Nasal Ala.

To evaluate the presence of cartilage in the lateral nasal ala (LNA) via histological analysis of alar Mohs surgical specimens. An accurate understanding of nasal anatomy is essential to optimize esthetic and reconstructive surgical outcomes. LNA anatomy is typically pictured to include discreet accessory and sesamoid cartilages; however, the authors debate the actual presence of these structures. A blinded reviewer analyzed 101 lateral nasal alar histologic specimens from 362 tissue blocks using a Hematoxylin and Eosin (H&E) stain to assess for the presence of cartilage. Out of the 362 tissue blocks histologically analyzed, only 1 included cartilaginous components. We conclude that the presence of sesamoid and accessory cartilages in the LNA within our sample set is exceedingly rare.

Molecular Characteristics of Staphylococcus aureus Strains Isolated from Nasal Cavity and Wound Infections Among Diabetic Patients.

Staphylococcus aureus is the most common pathogen contributing to diabetic foot infections (DFI). Nasal transmission of S. aureus potentially increases the risk of endogenous infection. The aim of this study was to determine the genetic diversity and antibiotic resistance profile of S. aureus isolates in nasal and wound samples from diabetic patients. A cross-sectional study was conducted from July 2018 to September 2019. S. aureus was isolated from the anterior nares and wounds of diabetic patients. All S. aureus isolates were characterized by detection of resistance and virulence genes (mecA, ermA, ermC, hla, hlb, hlg, sea, lukDE, pvl), staphylococcal cassette chromosome mec (SCCmec)-typing and staphylococcal protein A (spa)-typing. A total of 34 S. aureus were isolated from the wounds of 115 diabetic patients with DFI. Twenty-four S. aureus isolates were collected from the anterior nares of patients, and thirteen patients had concurrent S. aureus in nasal and wound specimens. The prevalence of methicillin-resistant S. aureus (MRSA) in nasal specimens was noticeable (41.7%), and the most common spa-type in nasal and wound specimens was t14870. Nearly half of the patients with concurrent S. aureus in wound and nasal specimens had similar isolates from both sites. Our data suggest that detection and screening of S. aureus colonization in the nasal cavity may prevent subsequent endogenous infections, particularly with MRSA strains.

Persistence of SARS-CoV-2 in saliva: Implications for late-stage diagnosis and infectious duration.

Saliva has been a COVID-19 diagnostic specimen of interest due to its simple collection, scalability, and yield. Yet COVID-19 testing and estimates of the infectious period remain largely based on nasopharyngeal and nasal swabs. We sought to evaluate whether saliva testing captured prolonged presence of SARS-CoV-2 and potential infectiousness later in the disease course. We conducted an observational study of symptomatic COVID-19 patients at University Hospital in Newark, NJ. Paired saliva and nasal specimens from 96 patients were analyzed, including longitudinal analysis of paired observations from 28 of these patients who had multiple time-points. Saliva detected significantly more cases of COVID-19 beyond 5 days (86.1% [99/115] saliva vs 48.7% [56/115] nasal, p-value < 0.001), 9 days (79.4% [50/63] saliva vs 36.5% [23/63] nasal, p-value < 0.001) and 14 days (71.4% [20/28] saliva vs 32.1% [9/28] nasal, p-value = 0.010) of symptoms. Additionally, saliva yielded lower cycle thresholds across all time periods, indicative of higher viral loads in saliva. In the longitudinal analysis, a log-rank analysis indicated that the survival curve for saliva was significantly different from the curve for nasal swabs (p<0.001) with a median survival time for saliva of 18 days compared to 13 days for nasal swabs. We additionally performed saliva viral cultures among a similar COVID-19 patient cohort and noted patients with positive saliva viral cultures between 7 to 28 days of symptoms. Findings from this study suggest that SARS-CoV-2 RNA persists longer and in higher abundance in saliva compared to nasal swabs, with potential of prolonged propagating virus. Testing saliva may thus increase yield for detecting potentially infectious virus even beyond the first five days of symptomatic COVID-19.

Clinical characteristics and outcomes of children with single or co-detected rhinovirus-associated acute respiratory infection in Middle Tennessee

BackgroundRhinovirus (RV) is one of the most common etiologic agents of acute respiratory infection (ARI), which is a leading cause of morbidity and mortality in young children. The clinical significance of RV co-detection with other respiratory viruses, including respiratory syncytial virus (RSV), remains unclear. We aimed to compare the clinical characteristics and outcomes of children with ARI-associated RV-only detection and those with RV co-detection—with an emphasis on RV/RSV co-detection.MethodsWe conducted a prospective viral surveillance study (11/2015–7/2016) in Nashville, Tennessee. Children < 18 years old who presented to the emergency department (ED) or were hospitalized with fever and/or respiratory symptoms of < 14 days duration were eligible if they resided in one of nine counties in Middle Tennessee. Demographics and clinical characteristics were collected by parental interviews and medical chart abstractions. Nasal and/or throat specimens were collected and tested for RV, RSV, metapneumovirus, adenovirus, parainfluenza 1–4, and influenza A–C using reverse transcription quantitative polymerase chain reaction assays. We compared the clinical characteristics and outcomes of children with RV-only detection and those with RV co-detection using Pearson’s χ2 test for categorical variables and the two-sample t-test with unequal variances for continuous variables.ResultsOf 1250 children, 904 (72.3%) were virus-positive. RV was the most common virus (n = 406; 44.9%), followed by RSV (n = 207; 19.3%). Of 406 children with RV, 289 (71.2%) had RV-only detection, and 117 (28.8%) had RV co-detection. The most common virus co-detected with RV was RSV (n = 43; 36.8%). Children with RV co-detection were less likely than those with RV-only detection to be diagnosed with asthma or reactive airway disease both in the ED and in-hospital. We did not identify differences in hospitalization, intensive care unit admission, supplemental oxygen use, or length of stay between children with RV-only detection and those with RV co-detection.ConclusionWe found no evidence that RV co-detection was associated with poorer outcomes. However, the clinical significance of RV co-detection is heterogeneous and varies by virus pair and age group. Future studies of RV co-detection should incorporate analyses of RV/non-RV pairs and include age as a key covariate of RV contribution to clinical manifestations and infection outcomes.

Accurate prediction of serum antibody levels from noninvasive saliva/nasal samples.

The importance of easily accessible, noninvasive samples, such as saliva, to effectively monitor serum antibody levels has been underscored by the SARS-CoV-2 (COVID-19) pandemic. Although a correlation between saliva and serum antibody titers has been observed, the ability to predict serum antibody levels from measurements in saliva is not well established. Herein, the authors demonstrate that measurements of SARS-CoV-2 antibody levels in both saliva and nasal specimens can be used to accurately determine serum levels by utilizing endogenous total IgG as an internal calibrator. They postulate that this method can be extended to the measurement of many different antibody analytes, making it of high interest for antibody therapeutic drug monitoring applications.

The association between allergic rhinitis and airway dysfunction and nasal endothelial damage and oxidative stress.

Although lower airway hyperresponsiveness is present in approximately one in three patients with allergic rhinitis (AR), the underlying mechanism remains unclear. To evaluate nasal patency and pulmonary functions in AR independently of the presence of asthma and to investigate the relationships between these and nasal oxidative stress parameters and endothelial damage. Seventy adolescents with AR (AR group - 27 with asthma and 43 without asthma) and 30 healthy controls (HC group) were included in this prospective, cross-sectional study. Endocan and oxidative biomarkers [total oxidant status (TOS), total antioxidant status (TAS), and oxidative stress index (OSI)] in nasal lavage fluid specimens; peak nasal inspiratory flow (PNIF); fractional exhaled nitric oxide (FeNO), and impulse oscillometry (zR5, zR20, and R5-20 for resistance and zX5 and zX20 for reactance) were investigated. Nasal endocan, TOS, and OSI values were higher in the AR group and TAS in the HC group. There was no difference between AR groups with and without asthma in terms of nasal endocan and oxidative biomarkers. FeNO levels and airway resistance (zR5, zR20, and R5-20) were higher in the AR group than in the HC group. However, there was no difference between the groups in PNIF. X5 was higher among the AR without asthma than in the other groups. Correlation between OSI and R5-20 was observed in the AR group. In the linear regression model, (logged) OSI was significantly predicted (logged) R5-20. The airways of adolescents with AR without asthma were as much affected as those of the AR with asthma, and this effect was associated with nasal endothelial damage and an increase in oxidative stress.

Pericytes may facilitate SARS-CoV-2 entry into the nervous system

Objective: Although the neurological and olfactory symptoms of coronavirus disease 2019 have been identified, the neurotropic properties of the causative virus, severe acute respiratory syndrome-associated coronavirus 2 (SARS-CoV-2), remain unknown. We sought to identify the susceptible cell types and potential routes of SARS-CoV-2 entry into the central nervous system, olfactory system, and respiratory system. Methods: We collected single-cell RNA data from normal brain and nasal epithelium specimens, along with bronchial, tracheal, and lung specimens in public datasets. The susceptible cell types that express SARS-CoV-2 entry genes were identified using single-cell RNA sequencing and the expression of the key genes at protein levels was verified by immunohistochemistry. We compared the coexpression patterns of the entry receptor angiotensin-converting enzyme 2 (ACE2) and the spike protein priming enzyme transmembrane serine protease (TMPRSS)/cathepsin L among the specimens. Results: The SARS-CoV-2 entry receptor ACE2 and the spike protein priming enzyme TMPRSS/cathepsin L were coexpressed by pericytes in brain tissue; this coexpression was confirmed by immunohistochemistry. In the nasal epithelium, ciliated cells and sustentacular cells exhibited strong coexpression of ACE2 and TMPRSS. Neurons and glia in the brain and nasal epithelium did not exhibit coexpression of ACE2 and TMPRSS. However, coexpression was present in ciliated cells, vascular smooth muscle cells, and fibroblasts in tracheal tissue; ciliated cells and goblet cells in bronchial tissue; and alveolar epithelium type 1 cells, AT2 cells, and ciliated cells in lung tissue. Conclusion: Neurological symptoms in patients with coronavirus disease 2019 could be associated with SARS-CoV-2 invasion across the blood–brain barrier via pericytes. Additionally, SARS-CoV-2–induced olfactory disorders could be the result of localized cell damage in the nasal epithelium.

Pre-analytical considerations in the development of a prototype SARS-CoV-2 antigen ARCHITECT automated immunoassay.

To evaluate pre-analytical challenges related to high-volume central laboratory SARS-CoV-2 antigen testing with a prototype qualitative SARS-CoV-2 antigen immunoassay run on the automated Abbott ARCHITECT instrument. Contrived positive and negative specimens and de-identified nasal and nasopharyngeal specimens in transport media were used to evaluate specimen and reagent on-board stability, assay analytical performance and interference, and clinical performance. TCID50/mL values were similar for specimens in various transport media. Inactivated positive clinical specimens and viral lysate (USA-WA1/2020) were positive on the prototype immunoassay. Within-laboratory imprecision was≤0.10 SD (<1.00S/C) with a≤10% CV (≥1.00S/C). Assay reagents were stable on board the instrument for 14days. No high-dose hook effect was observed with a SARS-CoV-2 stock of Ct 13.0 (RLU>1.0×106). No interference was observed from mucin, whole blood, 12 drugs, and more than 20 cross-reactants. While specimen stability was limited at room temperature for specimens with or without viral inactivation, a single freeze/thaw cycle or long-term storage (>30days) at-20°C did not adversely impact specimen stability or assay performance. Specificity of the prototype SARS-CoV-2 antigen immunoassay was≥98.5% and sensitivity was≥89.5% across two ARCHITECT instruments. Assay sensitivity was inversely correlated with Ct and was similar to that reported for the Roche Elecsys® SARS-CoV-2 Ag immunoassay. The prototype SARS-CoV-2 antigen ARCHITECT immunoassay is sensitive and specific for detection of SARS-CoV-2 in nasal and nasopharyngeal specimens. Endogenous proteases in mucus may degrade the target antigen, which limits specimen storage and transport times and complicates assay workflow.

Changes in percentage of GATA3+ regulatory T cells and their pathogenic roles in allergic rhinitis

To investigate the changes in percentage of GATA3+ regulatory T (Treg) cells in patients with allergic rhinitis (AR) and mouse models. The nasal mucosa specimens were obtained from 6 AR patients and 6 control patients for detection of nasal mucosal inflammation. Peripheral blood mononuclear cells (PBMC) were collected from 12 AP patients and 12 control patients to determine the percentages of Treg cells and GATA3+ Treg cells. In a C57BL/6 mouse model of AR, the AR symptom score, peripheral blood OVA-sIgE level, and nasal mucosal inflammation were assessed, and the spleen of mice was collected for detecting the percentages of Treg cells and GATA3+ Treg cells and the expressions of Th2 cytokines. Compared with the control patients, AR patients showed significantly increased eosinophil infiltration and goblet cell proliferation in the nasal mucosa (P < 0.01) and decreased percentages of Treg cells and GATA3+ Treg cells (P < 0.05). The mouse models of AR also had more obvious allergic symptoms, significantly increased OVA-sIgE level in peripheral blood, eosinophil infiltration and goblet cell hyperplasia (P < 0.01), markedly lowered percentages of Treg cells and GATA3+ Treg cells in the spleen (P < 0.01), and increased expressions of IL-4, IL-6 and IL-10 (P < 0.05). The percentage of GATA3+ Treg cells is decreased in AR patients and mouse models. GATA3+ Treg cells possibly participate in Th2 cell immune response, both of which are involved in the occurrence and progression of AR, suggesting the potential of GATA3+ Treg cells as a new therapeutic target for AR.

Performance of a SARS-CoV-2 RT-PCR Assay with Non-Traditional Specimen Types

During the first two years of the coronavirus disease 2019 (COVID-19) pandemic, nasopharyngeal (NP) specimens were the gold standard for clinical diagnostic testing. As information about the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causing the pandemic continued to be shared, it was clear that the virus could be detected in other specimen types during an active infection. The University of Louisville Infectious Diseases Laboratory accepted non-traditional specimen types, most without a paired, positive NP result, for research purposes only to support local epidemiology efforts. A real-time reverse transcription-polymerase chain reaction (RT-PCR) assay originally validated for NP specimens was used for non-traditional specimen types using a variety of specimen preparation methods. Limit of detection (LOD) studies allowed for direct comparison between NP, sputum, and breast milk specimen types. The primary aim of the study was to determine whether SARS-CoV-2 RNA could be detected in different human specimen types. The results showed that the non-traditional specimens were not inherently inhibitory since SARS-CoV-2 RNA was detected in 36 (14.5%) out of 249 non-traditional specimens, and the limit of detection for SARS-CoV-2 in breast milk and sputum was the same as for NP specimens. SARS-CoV-2 was not detected in 15 breast milk specimens from mothers with positive SARS-CoV-2 NP results. In addition, a direct comparison study showed that NP specimens performed better than paired nasal specimens. In conclusion, by analyzing real-time RT-PCR test results for these non-traditional specimen types, two benefits were realized. Health care providers gained additional epidemiologic information (since information was not to be used for managing or treating patients), and the laboratory gathered important information about specimen types for which complete method validation studies could be pursued in the future.

A Mini-Review On Emerging Trends and Recent Advances in Analytical Methods for Covid-19: Analytical Chemistry Perspective

In recent times SARS-CoV-2 pandemic, public health experts have slated testing, tracking infected people, as well as tracing their contacts as an efficient method to reduce the viral spread. Numerous diagnostic approaches are reported for detecting the coronavirus in public health, clinical and research laboratories. Most tests detect the infection directly by detecting the viral RNA while other test may indirectly detect the host antibodies in the course of detecting the infection. A diagnostic test during the pandemic should aid in making proper clinical decision within a short time frame. In this review, we describe the various techniques involved in sample collection from nasal, throat and sputum specimens, standard and developed methods for diagnosis of covid 19 such as the polymerase chain reaction (RT-PCR), CRISPR-based assays and Serological and immunological assays. General information on selected electroactive antiviral Covid-19 drugs have been highlighted including: Favipiravir (FAV), Remdesivir (REM), Lopinavir + Ritonavir (LOP + RIT), Hydroxychloroquine (HCQ) and Chloroquine (CQ), Ribavirin (RIB) and Sofosbuvir. This review presents the major standard, commercial and designed methods for detecting SARS-CoV-2 and their analytical performance. Also, chromatographic techniques have been highlighted as a rapid examination for SARS-CoV-2 identification, and some recent advances and emerging trends in analytical methods for covid-19 diagnosis have been addressed.

Utility of HumanIn VitroData in Risk Assessments of Influenza A Virus Using the Ferret Model

As influenza A viruses (IAV) continue to cross species barriers and cause human infection, the establishment of risk assessment rubrics has improved pandemic preparedness efforts. In vivo pathogenicity and transmissibility evaluations in the ferret model represent a critical component of this work. As the relative contribution of in vitro experimentation to these rubrics has not been closely examined, we sought to evaluate to what extent viral titer measurements over the course of in vitro infections are predictive or correlates of nasal wash and tissue measurements for IAV infections in vivo. We compiled data from ferrets inoculated with an extensive panel of over 50 human and zoonotic IAV (inclusive of swine-origin and high- and low-pathogenicity avian influenza viruses associated with human infection) under a consistent protocol, with all viruses concurrently tested in a human bronchial epithelial cell line (Calu-3). Viral titers in ferret nasal wash specimens and nasal turbinate tissue correlated positively with peak titer in Calu-3 cells, whereas additional phenotypic and molecular determinants of influenza virus virulence and transmissibility in ferrets varied in their association with in vitro viral titer measurements. Mathematical modeling was used to estimate more generalizable key replication kinetic parameters from raw in vitro viral titers, revealing commonalities between viral infection progression in vivo and in vitro. Meta-analyses inclusive of IAV that display a diverse range of phenotypes in ferrets, interpreted with mathematical modeling of viral kinetic parameters, can provide critical information supporting a more rigorous and appropriate contextualization of in vitro experiments toward pandemic preparedness. IMPORTANCE Both in vitro and in vivo models are employed for assessing the pandemic potential of novel and emerging influenza A viruses in laboratory settings, but systematic examinations of how well viral titer measurements obtained in vitro align with results from in vivo experimentation are not frequently performed. We show that certain viral titer measurements following infection of a human bronchial epithelial cell line are positively correlated with viral titers in specimens collected from virus-inoculated ferrets and employ mathematical modeling to identify commonalities between viral infection progression between both models. These analyses provide a necessary first step in enhanced interpretation and incorporation of in vitro-derived data in risk assessment activities and highlight the utility of employing mathematical modeling approaches to more closely examine features of virus replication not identifiable by experimental studies alone.

Effects of Neutrophil and Eosinophil Extracellular Trap Formation on Refractoriness in Chronic Rhinosinusitis With Nasal Polyps

This study investigated the clinical implications of neutrophil extracellular trap (NET) formation (NETosis) and eosinophil extracellular trap (EET) formation (EETosis) regarding refractoriness in chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP). Nasal polyp specimens were obtained from 117 patients with CRSwNP who received endoscopic sinus surgery. Disease control status at postoperative 1 year was assessed. Refractory cases were defined as partly controlled or uncontrolled cases according to the EPOS 2020 guidelines. NETosis and EETosis were evaluated through immunofluorescence staining (citrullinated histone H3-human neutrophil elastase and citrullinated histone-galectin-10, respectively) followed by manual counting. The z-score of NET and EET counts was used to define the following four groups: low extracellular trap formation (ETosis), NETosis-predominant, EETosis-predominant, and high-ETosis. The refractory and non-refractory groups showed significant differences in the tissue eosinophil count (P = 0.005) and EET count (P = 0.029). The tissue neutrophil count and the NET/neutrophil ratio were significantly different between the refractory and non-refractory groups of patients with neutrophilic CRS (P = 0.045, 0.031, respectively). Refractoriness significantly differed among the low-ETosis (30.77%), NETosis-predominant (47.83%), EETosis-predominant (56.67%), and high-ETosis (83.33%) groups (P = 0.005). The results of this study suggest that tissue Eosinophilia and EETosis may play a prognostic role, primarily in CRSwNP and thattissue neutrophilia and NETosis can play as prognostic biomarkers in neutrophilic CRSwNP.

A perspective on the effectiveness of SARS-Cov-2 rapid testing kits

Patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) are generally diagnosed by rapid antigen test (RAT) and reverse transcription-polymerase chain reaction (RT-PCR) tests. However, there is a conflict that the RT-PCR test is more effective than the RAT. The primary objective of this perspective is to compare and outline the effectiveness of SARS-CoV-2 rapid testing kits available in the market with RT-PCR tests. Recently published systematic reviews and meta-analysis reports containing comparative studies of RAT and RT-PCR were selected. The analysis results revealed that RAT kits provide remarkable specificity and sensitivity in the early stages of infection, particularly when the viral load is huge, compared to RT-PCR. In addition, a trustworthy substitute for nasopharyngeal sampling is the use of nasal specimens for antigen detection, which are patient-friendly and somewhat sensitive. RAT could be beneficial in the combat against the COVID-19 pandemic, but it has to be combined with appropriate monitoring of results that come back negative.

Molecular epidemiology of clinical filamentous fungi in Qatar beyond Aspergillus and Fusarium with notes on the rare species

Due to an increasing number of patients at risk (i.e., those with a highly compromised immune system and/or receiving aggressive chemotherapy treatment), invasive fungal infections (IFI) are increasingly being reported and associated with high mortality rates. Aspergillus spp., particularly A. fumigatus, is the major cause of IFI caused by filamentous fungi around the world followed by Fusarium spp., however, other fungi are emerging as human pathogens. The aim of this study was to explore the epidemiology and prevalence of the non-Aspergillus and non-Fusarium filamentous fungi in human clinical samples over an 11-year period in Qatar using molecular techniques. We recovered 53 filamentous fungal isolates from patients with various clinical conditions. Most patients were males (75.5%), 9.4% were immunocompromised, 20.7% had IFI, and 11.3% died within 30 days of diagnosis. The fungal isolates were recovered from a variety of clinical samples, including the nasal cavity, wounds, respiratory samples, body fluids, eye, ear, tissue, abscess, and blood specimens. Among the fungi isolated, 49% were dematiaceous fungi, followed by Mucorales (30%), with the latter group Mucorales being the major cause of IFI (5/11, 45.5%). The current study highlights the epidemiology and spectrum of filamentous fungal genera, other than Aspergillus and Fusarium, recovered from human clinical samples in Qatar, excluding superficial infections, which can aid in the surveillance of uncommon and emerging mycoses.