scholarly journals 729. Improved Detection of Adenovirus with the FilmArray Respiratory Panel 2 Panel in a Pediatric Population

2018 ◽  
Vol 5 (suppl_1) ◽  
pp. S262-S262
Author(s):  
Kathy Everhart ◽  
Huanyu Wang ◽  
Li Xu ◽  
Amy Leber
Keyword(s):  
Respiratory Infections ◽  
Pediatric Population ◽  
Human Adenovirus ◽  
Fold Increase ◽  
Clinical Samples ◽  
Research Support ◽  
Respiratory Illnesses ◽  
Two Samples ◽  
Type C ◽  
Hospital Research

Abstract Background Human adenovirus (ADV) respiratory infections are associated with up to 8% of all identified viral causes of acute respiratory illnesses, especially among young children. They are therefore included as part of multiplex syndromic respiratory testing. Earlier versions of the FilmArray Respiratory Panel were targeted to genogroups B, C, and E. However, all adenovirus genogroups are associated with disease particularly in immunocompromised patients. Recently, a new version of the FilmArray Respiratory Panel 2 (RP2) has been FDA cleared with significant modifications to the adenovirus assay. The goal of this study was to compare the RP2 adenovirus assay to that of the previous version (RP1.7) and our laboratory-developed (LD) adenovirus PCR. Methods Analytical comparison: Ten stocks of know adenovirus serotypes representing genogroups A-F were diluted in M4 media and tested on RP2, RP1.7 and LD PCR to determine the relative limits of detection (LOD). Clinical comparison: A total of 423 pediatric nasopharyngeal samples were tested using RP2, RP1.7, adenovirus LD PCR. In addition we performed genotyping PCRs on most adenovirus positive samples based on the availability. Results Analytical evaluation revealed that for the 10 serotypes (18, 32, 7, 14, 5, 6, 20, 29, 4 and 40), RP2 showed at least 100-fold increase in sensitivity for six serotypes representing genogroups A, D and F. For B, C and E, the relative LODs were comparable. In the 423 clinical samples, there was an overall agreement of 94% between RP2 and RP1.7. Among those RP2+/RP1.7− samples (n = 20), 17 samples were confirmed ADV positive by LD PCR. They all had low viral burden (Ct values >30), among them nine samples had sole detections of adenovirus types A, D and F. Five samples were RP2-/RP1.7+, two samples were confirmed by LD PCR and both were type C. Overall, there was a 22% increase in Adenovirus with RP2 compared with RP1.7. Conclusion The RP2 adenovirus assay has enhanced inclusivity and lower LOD for all adenovirus genogroups in comparison to RP1.7. Coupled with its faster run time and additional targets, RP2 represents a significant improvement for the syndromic detection of respiratory infections. Disclosures A. Leber, Nationwide Children’s Hospital: Research Contractor, Research support.

Identification of adenovirus species in Iranian pediatric population with severe acute respiratory infections

2019 ◽  
Vol 14 (9) ◽  
pp. 577-583
Author(s):  
Nazanin-Zahra Shafiei-Jandaghi ◽  
Jila Yavarian ◽  
Somayeh Shatizadeh Malekshahi ◽  
Maryam Naseri ◽  
Azadeh Shadab ◽  
...  
Keyword(s):  
Phylogenetic Tree ◽  
Molecular Epidemiology ◽  
Nested Pcr ◽  
Acute Respiratory Infection ◽  
Respiratory Infections ◽  
Pediatric Population ◽  
Human Adenovirus ◽  
Acute Respiratory Infections ◽  
Severe Acute Respiratory Infection ◽  
Severe Acute Respiratory Infections

Aim: In this study we aimed to find the rate of human adenovirus (HAdV) infections and identify HAdVs molecular epidemiology in children younger than 5 years old with clinical diagnosis of severe acute respiratory infections in Iran. Patients & methods: A semi-nested PCR with in-house designed primers for HAdV was performed on 200 oropharyngeal swabs collected within 5 days from the onset of symptoms. The positive samples were subjected to sequencing and a phylogenetic tree was drawn. Results: Out of 200 specimens, 71 (35.5%) cases were positive for HAdVs. Detected strains matched with HAdV species B (80.3%), C (15.5%), D (2.8%) and E (1.8%). HAdV- B14/55 and HAdV-B3 were the most prevalent types, respectively. Conclusion: This study showed that infections with HAdV species B, were common in children, who were clinically diagnosed as severe acute respiratory infection cases in Iran. The results could be useful for future epidemiological researches.

scholarly journals Nasopharyngeal colonization with pathobionts is associated with susceptibility to respiratory illnesses in young children

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0243942
Author(s):  
Timothy J. Chapman ◽  
Matthew C. Morris ◽  
Lei Xu ◽  
Michael E. Pichichero
Keyword(s):  
Clustering Analysis ◽  
Moraxella Catarrhalis ◽  
Respiratory Infections ◽  
Bacterial Colonization ◽  
Clinical Samples ◽  
Early Age ◽  
Nasopharyngeal Colonization ◽  
Streptococcus Pneumonia ◽  
Respiratory Illnesses ◽  
Microbiome Diversity

Some children are more susceptible to viral and bacterial respiratory infections in the first few years of life than others. However, the factors contributing to this susceptibility are incompletely understood. In a retrospective analysis of clinical samples collected from a prospectively-enrolled cohort of 358 children we sought associations between physician-attended illness visits and bacterial colonization in the first five years of life. A subset of children was identified by unsupervised clustering analysis as infection and allergy prone (IAP). Several respiratory infection- and allergy-mediated illnesses co-occurred at higher rates in IAP children, while the rates of other illnesses were not significantly different between the groups. Analyses of nasopharyngeal (NP) pathobionts and microbiota commensals showed that early age of first colonization with pathobionts Streptococcus pneumonia, non-typeable Haemophilus influenzae, and Moraxella catarrhalis was associated with IAP children, and particularly Moraxella abundance was negatively associated with NP microbiome diversity. We conclude that mucosal pathobiont exposures in early life can influence susceptibility to respiratory illnesses in children.

scholarly journals 237. Adenovirus Types in Children with Acute Respiratory Illnesses in Nashville Over Two Respiratory Seasons

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S135-S136
Author(s):  
Varvara Probst ◽  
Andrew Speaker ◽  
Laura S Stewart ◽  
Claudia Guevara ◽  
Susan I Gerber ◽  
...  
Keyword(s):  
Real Time ◽  
Pediatric Population ◽  
Human Adenovirus ◽  
Respiratory Viruses ◽  
Early Spring ◽  
Hospital Inpatient ◽  
Respiratory Illnesses ◽  
Chart Abstraction ◽  
Co Detection ◽  
Nose And Throat

Abstract Background Human adenovirus (HAdV) types 1–7, 11, 14, 16, and 21 within species B, C, and E are commonly associated with acute respiratory illnesses (ARI) in children. We sought to compare demographics, clinical characteristics, and outcomes of HAdV types with children who presented with fever and/or respiratory symptoms. Methods Children < 18 years with fever and/or ARI seen at Vanderbilt Children’s Hospital inpatient and emergency department settings from December of 2016 to October of 2018 were enrolled. Interviews and chart abstraction were conducted. Mid-turbinate nose and throat swab specimens were collected and tested by real-time RT-PCR for common respiratory viruses including HAdV. HAdV molecular typing was performed by type-specific real-time PCR assays for types 1–7, 11, 14, 16, and 21 targeting the hexon gene using published methods. Results Of 5111 ARI cases, 206 (4%) were HAdV-positive with a median age of 16 months (IQR 9–30); 57% male, 47% White, 40% Black, 33% Hispanic, 20% admitted, and 24% of hospitalized required oxygen support. Of the 206, 186 (90%) were able to be typed with more than one types detected in 13 (7%) cases. Distribution of HAdV types among single detections (n = 173) is shown in Figure 1; HAdV-1 and HAdV-2 were most common. Children with HAdV-2 were younger (median age 12 months vs. 15 months (HAdV-1) and 59 months (all other types), P < 0.001), and those with HAdV-1 were less likely to be male (44% vs. 65% for both HAdV-2 and other types, P = 0.029). Figure 2 displays HAdV detections over time, with winter and early spring peaks. Co-detection with other respiratory viruses occurred in 47% of cases; the most common among typable HAdV were rhinovirus/enterovirus in 30/186 (16%) and RSV in 19/186 (10%). Distribution among HAdV types is shown in Figure 3. Conclusion HAdV-1 and HAdV-2 were more prevalent than other HAdV types over two respiratory seasons in the Nashville area with peak cases in December-March. Children with HAdV-1 and HAdV-2 had some demographic differences. Further studies with a larger sample size for HAdV typing are needed in the pediatric population to determine whether additional clinically-relevant differences between HAdV types exist. Disclosures All authors: No reported disclosures.

Detection of viruses by electron microscopy: Increased sensitivity by direct micro-ultracentrifugation of samples

1987 ◽  
Vol 45 ◽  
pp. 908-909
Author(s):  
Alain R. Trudel ◽  
M. Trudel
Keyword(s):  
Electron Microscopy ◽  
Fold Increase ◽  
Observation Time ◽  
Clinical Samples ◽  
Maximum Speed ◽  
Viral Particles ◽  
Structural Details ◽  
Latex Spheres ◽  
Increased Sensitivity ◽  
Size Of Particles

AirfugeR (Beckman) direct ultracentrifugation of viral samples on electron microscopy grids offers a rapid way to concentrate viral particles or subunits and facilitate their detection and study. Using the A-100 fixed angle rotor (30°) with a K factor of 19 at maximum speed (95 000 rpm), samples up to 240 μl can be prepared for electron microscopy observation in a few minutes: observation time is decreased and structural details are highlighted. Using latex spheres to calculate the increase in sensitivity compared to the inverted drop procedure, we obtained a 10 to 40 fold increase in sensitivity depending on the size of particles. This technique also permits quantification of viral particles in samples if an aliquot is mixed with latex spheres of known concentration.Direct ultracentrifugation for electron microscopy can be performed on laboratory samples such as gradient or column fractions, infected cell supernatant, or on clinical samples such as urine, tears, cephalo-rachidian liquid, etc..

scholarly journals 1718. Respiratory Syncytial Virus (RSV) Surveillance in the Department of Veterans Affairs (VA), 2010-2018

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S843-S843
Author(s):  
Cynthia Lucero-Obusan ◽  
Patricia Schirmer ◽  
Gina Oda ◽  
Mark Holodniy
Keyword(s):  
Respiratory Syncytial Virus ◽  
Respiratory Infections ◽  
Diagnostic Testing ◽  
Fold Increase ◽  
Respiratory Illness ◽  
Veterans Affairs ◽  
Department Of Veterans Affairs ◽  
Management And Development ◽  
Syncytial Virus ◽  
National Trends

Abstract Background Respiratory Syncytial Virus (RSV) is an increasingly recognized cause of acute respiratory illness in older adults, leading to an estimated 177,000 hospitalizations and 14,000 deaths each year in the US. In adult populations, diagnostic testing for RSV has historically been underutilized. Herein, we examine national trends in RSV testing and infection across the Veterans Affairs (VA) healthcare system. Methods Electronic RSV laboratory testing results, ICD-coded hospitalizations and outpatient encounters were obtained from VA’s Praedico Surveillance System (1/1/2010-12/31/2018). Patients were reviewed for positive results, repeat testing, and demographics. Antibody tests were excluded. Results A total of 102,251 RSV results were included. Overall, 4,372 (4.3%) specimens from 4,263 unique individuals were positive with a median age of 67 years (range 0-101) and 90% were male. 1,511 individuals (35.4%) also had an RSV-coded hospitalization. RSV type was specified for only 7.8% of positives (Table). During 2010-2018 there were 2,522 RSV-coded hospitalizations (median length of stay = 4 days) among 2,444 unique individuals, which included 413 ICU stays (16.4%) and 98 deaths (3.9%) during the RSV-coded hospitalization. Approximately 78% of RSV-coded hospitalizations within VA (excluding all non-VA hospitalizations) had a documented positive test result. A greater than 15-fold increase in RSV tests performed, hospitalizations and outpatient encounters was observed from 2010-2018, although the percent testing positive remained relatively stable (Figure, Table). Figure. Testing for Respiratory Syncytial Virus (RSV), Department of Veterans Affairs, 2010-2018. Table. Select RSV Surveillance Metrics, Department of Veterans Affairs, 2010-2018 Conclusion RSV testing and identification of patients with RSV infection increased dramatically during the time period analyzed, likely due to increased availability of PCR-based multi-pathogen panels and duplex assays. While the percentage of tests positive for RSV remained relatively stable, the rise in coded hospitalizations may be due to increased testing for RSV among hospitalized Veterans with severe respiratory infections. These surveillance data may allow for further characterization of RSV disease burden estimates which can help inform clinical management and development of interventions for adults, such as vaccines and antiviral therapies. Disclosures All Authors: No reported disclosures

scholarly journals 1186. Cefazolin inoculum effect predicts reduced susceptibility to other antibiotics and patient outcomes in MSSA endovascular infections

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S617-S617
Author(s):  
Dan Smelter ◽  
Sue McCrone ◽  
Warren Rose
Keyword(s):  
Infective Endocarditis ◽  
Mortality Rate ◽  
Patient Outcomes ◽  
Treatment Options ◽  
Fold Increase ◽  
Specific Mechanism ◽  
Research Support ◽  
High Burden ◽  
Low Toxicity ◽  
Inoculum Effect

Abstract Background MSSA Infective endocarditis (IE) is inherently a high-burden infection with up to a 30% mortality rate. Cefazolin is an appealing treatment option for IE with low toxicity and a favorable dosing scheme. However, cefazolin has been associated with treatment failure in IE, attributed to an inoculum effect. The specific mechanism underlying the cefazolin inoculum effect (CIE) remains undetermined, but CIE has been linked to both blaZ expression and agr dysfunction. This study aims to determine whether CIE is linked to reduced susceptibility to other antibiotics and worse outcomes regardless of therapy in MSSA endovascular infections. Methods Sixty-four MSSA strains were collected from patients with endovascular infections not treated with cefazolin. To determine CIE phenotype, strains were cultured and MICs assayed for cefazolin, nafcillin, and vancomycin at 107 CFU/mL for high-inocula (HI) and 105 CFU/mL for standard-inocula (SI). This study defined CIE as a ≥ 4-fold increase in MIC at HI compared to SI, with at least an MIC of 4 mg/L at HI. Nitrocefin disks identified blaZ expression, and beta lysin disks were used to determine hemolysin type and agr function. Patient outcomes of mortality and bacteremia duration were assessed across cohorts. Results Twenty-four strains exhibit a CIE (38%), with 10 strains having an MIC of ≥ 32mg/L at HI. Nafcillin and vancomycin also had an inoculum effect, uncoupled from the CIE and occurring at a lower frequency and amplitude at HI. Presence of CIE had a greater association with blaZ expression (71% vs 25%) than agr dysfunction (38% vs 20%). 50% (9/18) of CIE infections were cleared within 48 hours while 77% (20/26) of CIE-negative infections were cleared within 48 hours (P=0.106). However, presence of CIE was not associated with increased mortality (25% CIE-positive vs 35%; P=0.578) Conclusion Previous studies for CIE failed to enrich for isolates from endovascular sources, where inocula are known to be high. This study presents one of the largest endovascular source cohorts for CIE evaluation. It identifies that CIE prevalence (38%) is higher than reports from diverse infection sources (10-36%). CIE appears to predict bacteremia duration with other MSSA treatment options, suggesting mechanisms independent of blaZ and agr function for this phenomenon. Disclosures Warren Rose, PharmD, MPH, Merck (Grant/Research Support)Paratek (Grant/Research Support)

scholarly journals Discovery of a Small Molecule Inhibitor of Human Adenovirus Capable of Preventing Escape from the Endosome

2021 ◽  
Vol 22 (4) ◽  
pp. 1617
Author(s):  
Jimin Xu ◽  
Judith Berastegui-Cabrera ◽  
Marta Carretero-Ledesma ◽  
Haiying Chen ◽  
Yu Xue ◽  
...  
Keyword(s):  
Potent Inhibitor ◽  
Antiviral Drug ◽  
Human Adenovirus ◽  
Yield Reduction ◽  
Respiratory Illnesses ◽  
Molecule Inhibitor ◽  
Wide Range ◽  
Ic50 Values ◽  
Life Threatening ◽  
Salicylamide Derivatives

Human adenoviruses (HAdVs) display a wide range of tissue tropism and can cause an array of symptoms from mild respiratory illnesses to disseminated and life-threatening infections in immunocompromised individuals. However, no antiviral drug has been approved specifically for the treatment of HAdV infections. Herein, we report our continued efforts to optimize salicylamide derivatives and discover compound 16 (JMX0493) as a potent inhibitor of HAdV infection. Compound 16 displays submicromolar IC50 values, a higher selectivity index (SI > 100) and 2.5-fold virus yield reduction compared to our hit compound niclosamide. Moreover, unlike niclosamide, our mechanistic studies suggest that the antiviral activity of compound 16 against HAdV is achieved through the inhibition of viral particle escape from the endosome, which bars subsequent uncoating and the presentation of lytic protein VI.

scholarly journals Human adenovirus species in children with acute respiratory illnesses

2021 ◽  
Vol 134 ◽  
pp. 104716
Author(s):  
Varvara Probst ◽  
Emily K. Datyner ◽  
Zaid Haddadin ◽  
Danielle A. Rankin ◽  
Lubna Hamdan ◽  
...  
Keyword(s):  
Human Adenovirus ◽  
Respiratory Illnesses

Distinct Patterns of Emergence of SARS-CoV-2 Spike Variants including N501Y in Clinical Samples in Columbus Ohio

2021 ◽  
Author(s):  
Huolin Tu ◽  
Matthew R Avenarius ◽  
Laura Kubatko ◽  
Matthew Hunt ◽  
Xiaokang Pan ◽  
...  
Keyword(s):  
Clinical Impact ◽  
Clinical Samples ◽  
Membrane Glycoprotein ◽  
Receptor Binding Domain ◽  
Furin Cleavage ◽  
S Gene ◽  
The United Kingdom ◽  
Furin Cleavage Site ◽  
Two Samples ◽  
The Uk

AbstractFollowing the worldwide emergence of the p.Asp614Gly shift in the Spike (S) gene of SARS-CoV-2, there have been few recurring pathogenic shifts occurring during 2020, as assessed by genomic sequencing. This situation has evolved in the last several months with the emergence of several distinct variants (first identified in the United Kingdom and South Africa) that manifest multiple changes in the S gene, particularly p.Asn501Tyr (N501Y), that likely have clinical impact. We report here the emergence in Columbus, Ohio in December 2020 of two novel SARS-CoV-2 clade 20G variants. One variant, that has become the predominant virus found in nasopharyngeal swabs in the December 2020-January 2021 period, harbors S p.Gln677His (Q677H), affecting a consensus QTQTN domain near the S1/S2 furin cleavage site, nucleocapsid (N) p.Asp377Tyr (D377Y) and membrane glycoprotein (M) p.Ala85Ser (A85S) mutations, with additional S mutations in subsets. The other variant present in two samples, contains S N501Y, which is a marker of the UK-B.1.1.7 (clade 20I/501Y.V1) strain, but lacks all other mutations from that virus. The Ohio variant is from a different clade and shares multiple mutations with the clade 20G viruses circulating in the area prior to December 2020. These two SARS-CoV-2 viruses, which we show are also present and evolving currently in several other parts of North America, add to the diversity of S gene shifts occurring worldwide. These and other shifts in this period of the pandemic support multiple independent acquisition of functionally significant and potentially complementing mutations affecting the S QTQTN site (Q675H or Q677H) and certain receptor binding domain mutations (e.g., E484K and N501Y).

scholarly journals Clinico-epidemiological profile of acute respiratory infections and malnutrition in urban and rural population of central India

2016 ◽  
Vol 4 (1) ◽  
pp. 159
Author(s):  
Pranav G. Jawade ◽  
Neelam D. Sukhsohale ◽  
Gayatri G. Jawade ◽  
Binish Z. A. Khan ◽  
Pratik K. Kakani ◽  
...  
Keyword(s):  
Rural Population ◽  
Urban Population ◽  
Respiratory Infections ◽  
Severe Acute Malnutrition ◽  
Pediatric Population ◽  
Central India ◽  
Developing Nations ◽  
Acute Malnutrition ◽  
Acute Respiratory Infections ◽  
Cross Sectional

Background: Acute respiratory infections (ARI) and Malnutrition in children have tremendous burden on the health care sector of developing nations including India. The intensity with which these conditions are holding grip in the community is indeed a matter of concern and hence developing nations should develop an insight to assess the severity of it has become a necessity.Methods: A hospital based cross sectional study was carried out in children aged 0-14 years. Children were clinically assessed and diagnosis was made as URTI or LRTI Also anthropometry was performed and accordingly children were divided into categories of no malnutrition (NM), severe acute malnutrition (SAM) and moderate acute malnutrition (MAM) in ‘under 5’ years age children according to WHO guidelines, whereas children aged ‘above 5’ years were categorized as per the IAP guidelines.Results: It was observed that most of the mothers of children were illiterate with inadequate or absent ventilation and use of biomass fuels (chulha) for cooking purpose in households of rural children. The proportion of malnutrition was found to be equal in Under 5 children. In ‘above 5 years’ age study subjects, 52.4% of urban study subjects were normal as compared to 16.7% of rural study subjects.Conclusions: Our study implies that ARI and Malnutrition definitely is more prevalent in the pediatric population. Prevalence of URTI was found to be on a higher side in the rural population and LRTI prevalence was found to be higher in the urban population. Similarly, the prevalence of malnutrition was almost similar in ‘Under 5’ aged children, whereas the prevalence of malnutrition in ‘Above 5’ aged children was higher in the rural population than urban population.

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