Parainfluenza Research Articles

2652. Detection of respiratory viruses by air sampling and student and staff surveillance testing in two elementary schools

Abstract Background The COVID-19 pandemic highlighted the need for better data to understand how ventilation may impact respiratory virus transmission. We evaluated if there was an association between respiratory viruses detected in school room air samples and nasal swabs from students/ staff in the same room. Methods We performed air and respiratory sampling to detect respiratory viruses at 3 time points between Feb 27 - Mar 28, 2023 in 2 elementary schools. Three classrooms, a multipurpose room, and school nurse’s office in each school had 1 air sample/ room collected using Thermo Scientific AerosolSense™ sampler during 8 school hours. Concomitant respiratory viral testing, using self-collected nasal swabs, was performed in students/ staff enrolled in a respiratory virus surveillance school testing program. Air and respiratory specimens were tested for adenovirus (AdV); coronaviruses (CoV) 229E, HKU1, NL63, OC43; human metapneumovirus (hMPV), influenza A and B (Flu), parainfluenza virus 1-4 (PIV), respiratory syncytial virus (RSV), rhinovirus/ enterovirus (RV/EV), and SARS-CoV-2 using the Hologic® Panther Fusion® Respiratory Assay. Detected respiratory viruses in room air and respiratory samples were compared to determine likelihood of capturing current human respiratory viral infections in room air and additional viruses circulating from unsampled students/ staff in the same room. RV and AdV in human respiratory samples were typed by partial genomic sequencing to understand if viruses were similar. Results Ten rooms, including 6 classrooms with ≈25 people/ room, were sampled at 3 occasions. At least 1 respiratory virus was detected in 27/ 30 (90%) air samples. 59 samples were obtained from 20 students and 5 staff in these rooms; 24 (41%) were positive for ≥1 virus and 11/24 (46%) rooms with concomitant air and respiratory samples had the same viruses detected (Table). RV typing demonstrates that both participants from Room E, Round 1 and 2 had RV-B27. Both participants from Room F, Round 2 with single AdV infections had AdV B3.Table.Concomitant respiratory virus detection from student and staff nasal swabs and air samplers (N=student/staff tested per room) Conclusion Concomitant respiratory virus surveillance in air samples and students/ staff can be performed to better understand the role of ventilation and respiratory virus transmission. Additional studies, including sequencing data and symptom assessment, are needed. Disclosures Rangaraj Selvarangan, BVSc, PhD, D(ABMM), FIDSA, FAAM, Abbott: Honoraria|Altona Diagnostics: Grant/Research Support|Baebies Inc: Advisor/Consultant|BioMerieux: Advisor/Consultant|BioMerieux: Grant/Research Support|Bio-Rad: Grant/Research Support|Cepheid: Grant/Research Support|GSK: Advisor/Consultant|Hologic: Grant/Research Support|Lab Simply: Advisor/Consultant|Luminex: Grant/Research Support

2653. Duration of Respiratory Virus Detection by Multipathogen PCR Testing after Acute Respiratory Illness

Abstract Background Persons with respiratory viral infections (RVI) may shed detectable virus after symptom recovery. Duration of detection by molecular methods is not well described for non-SARS-CoV-2 respiratory viruses. Methods School KIDS is a prospective respiratory viral surveillance program in a Missouri school district. Participating students and staff with RVI symptoms can receive respiratory viral testing using self-collected nasal swabs. Symptomatic participants with detectable virus(es) on primary swabs were scheduled to receive 2 convalescent swabs weekly after initial test during weekly school surveillance testing. All convalescent tests performed within 21 days of the primary specimen were included. Only participants providing ≥ 1 convalescent swab were included. Specimens were tested using Hologic® Panther Fusion® or QIAstat-Dx PCR assays that included adenovirus (AdV); seasonal coronaviruses (sCoV) 229E, HKU1, NL63, OC43; human metapneumovirus, influenza A and B (Flu), parainfluenza viruses 1-4, respiratory syncytial virus, rhinovirus/ enterovirus (RV/EV), and SARS-CoV-2. We used survival analysis to quantify median number of days until individuals tested negative for each virus. Results From November 3, 2022 – April 14, 2023, 344 symptomatic participants (141 preK/elem, 49 middle, 22 high, 132 staff) submitted 586 primary specimens; 292 (50%) were positive for a virus. For positive primary specimens, median [IQR] time of symptom onset to specimen collection was 2 [1,3] days. A total of 320 viruses were detected; most common were RV/EV (37%) and sCoVs (25%) (Table). An additional 370 convalescent specimens were collected (87 specimens on days 1-7 following primary positive; 166 specimens on days 8-14; and 117 specimens on days 15-21). Median number of days [95% confidence interval] when repeat convalescent tests became negative varied by virus (AdV 15 [11,undefined] days, Flu A 14 [9,19] days, RV/EV 12 [11,14] days, SARS-CoV-2 14 [14,17] days, sCoV 10 [9,12] days) (Figure).Table.Characteristics of primary positive and convalescent testing (i.e. ongoing positivity) of nasal swabs from preK—12 grade students and staff with respiratory symptoms—Missouri, November 2022 -April 2023Figure.Cumulative probability of testing negative for a respiratory virus on subsequent respiratory viral testing Conclusion Students and staff continued to have detectable virus by nasal swabs 10-15 days after symptomatic positive specimen. Further data are needed to determine when participants are no longer infectious. Disclosures Rangaraj Selvarangan, BVSc, PhD, D(ABMM), FIDSA, FAAM, Abbott: Honoraria|Altona Diagnostics: Grant/Research Support|Baebies Inc: Advisor/Consultant|BioMerieux: Advisor/Consultant|BioMerieux: Grant/Research Support|Bio-Rad: Grant/Research Support|Cepheid: Grant/Research Support|GSK: Advisor/Consultant|Hologic: Grant/Research Support|Lab Simply: Advisor/Consultant|Luminex: Grant/Research Support

2618. Four Respiratory Viruses are Involved in the Majority of Community-Acquired Alveolar Pneumonia (CAAP) Episodes in Children < 5 Years

Abstract Background Community-acquired alveolar pneumonia (CAAP) is considered a bacterial disease in most cases. However, multiple studies in young children have shown a high correlation of its incidence with the epidemiological activity of four respiratory viruses (RSV, hMPV, influenza, and parainfluenza [pneumonia-associated viruses; PAV]). This strongly suggests an important role for bacterial-viral coinfections. The current study examines the detection rate of PAV in radiologically-confirmed CAAP in young children. Methods Hospitalization for CAAP episodes and viral detection were prospectively studied among all children under 5 years living in southern Israel (the Negev district); over 95% of these children are born and treated at the only medical center in the region. This prospective active surveillance was previously described (Dagan, EBioMedicine, 90:104493, 2023). Viral activity was defined by all PAV-positive tests in children under 5 years in the community. Nasal samples of children with CAAP were obtained for virus detection by PCR within 48 hours from admission. Only episodes tested for all four PAVs were included. The monthly incidence of PAV-positive CAAP was calculated by extrapolating to all-CAAP episodes, in each month. Results During the study period, 2,592 CAAP hospitalizations occurred. Specimens for all four PAVs were obtained in 1,851 (71.4%) CAAP episodes (< 12 months, 1,009; 12-23 months, 443; 24-59 months, 399) (Table). Overall, 59% of all episodes were positive for one or more PAVs, 64% of which were RSV (either as single or mixed detection). The surveillance of PAV activity in the community yielded 9,021 positive specimens for one or more PAVs. The dynamic patterns of both all-cause and PAV-positive CAAP hospitalizations from July 2016 through December 2022 closely resembled that of the PAV activity in the community, including the unusual pattern during the COVID-19 years 2020 through 2022 (Figure 1). The PAV-CAAP episodes were the main contributors of the CAAP seasonality (Figure 2). Conclusion RSV, followed by hMPV, and to a lesser degree influenza and parainfluenza viruses, were the main viruses detected in children under 5 years hospitalized for CAAP. These four viruses were responsible for the seasonal pattern of CAAP hospitalizations. Disclosures Ron Dagan, Professor MD, GSK: Honoraria|MedIMmune/AstraZeneca: Grant/Research Support|MSD: Advisor/Consultant|MSD: Grant/Research Support|MSD: Honoraria|Pfizer: Advisor/Consultant|Pfizer: Expert Testimony|Pfizer: Grant/Research Support|Pfizer: Honoraria|Sanofi Pasteur: Honoraria David Greenberg, Professor MD, GSK: Advisor/Consultant|GSK: Honoraria|MSD: Advisor/Consultant|MSD: Grant/Research Support|MSD: Honoraria|Pfizer: Advisor/Consultant|Pfizer: Honoraria Shalom Ben-Shimol, Dr. MD, GSK: Honoraria|MSD: Advisor/Consultant|MSD: Honoraria|Pfizer: Advisor/Consultant|Pfizer: Grant/Research Support|Pfizer: Honoraria Daniel M. Weinberger, PhD, GSK: Advisor/Consultant|Merck: Advisor/Consultant|Merck: Grant/Research Support|Pfizer: Advisor/Consultant|Pfizer: Grant/Research Support Dana Danino, Dr. MD, Pfizer: Grant/Research Support

2655. Phase 1 Safety and Immunogenicity Results of Two Investigational mRNA Vaccines, mRNA-1345, a Respiratory Syncytial Virus Vaccine, and mRNA-1653, a Human Metapneumovirus and Parainfluenza Virus Type 3 Combination Vaccine in Seropositive Young Children

Abstract Background Respiratory syncytial virus (RSV), human metapneumovirus (hMPV), and parainfluenza virus type 3 (PIV3) are common respiratory illnesses in children. Two investigational vaccines, mRNA-1345, encoding the RSV prefusion stabilized F (preF) glycoprotein, and mRNA-1653, encoding the hMPV and PIV3 F glycoproteins, are in clinical trials. Methods Two phase 1, randomized, observer-blind, placebo-controlled trials in children aged 12-59 months assessed safety and immunogenicity of mRNA-1345 (NCT04528719) and mRNA-1653 (NCT04144348). In the mRNA-1345 trial, RSV-seropositive children (N=46) were randomized to receive 3 doses of mRNA-1345 (15 µg or 30 µg) or placebo 2 months apart. In the mRNA-1653 trial, hMPV- and PIV3-seropositive children (N=27) were randomized to receive 2 doses of mRNA-1653 (10 µg or 30 µg) or placebo 2 months apart. Interim data through Month (M) 5 for mRNA-1345 and M3 for mRNA-1653 are reported. Results mRNA-1345 and mRNA-1653 were well-tolerated. The most frequently reported solicited local adverse reaction (AR) was tenderness at injection site (mRNA-1345, 35.7%-71.4%; placebo, 26.7%-42.9% and mRNA-1653, 44.4%-60.0%; placebo, 12.5%-30.0%); solicited systemic ARs (mRNA-1345, 12.5%-53.3%; placebo, 33.3%-50.0% and mRNA-1653, 33.3%-55.6%; placebo, 12.5%-60.0%) were mostly grade 1/2. One mRNA-1345 injection boosted RSV neutralizing antibody (nAb) titers (geometric mean fold rise [GMFR] over baseline: RSV-A=18.9-34.9; RSV-B=7.2-14.3) and RSV preF and postF binding antibody (bAb) concentrations (GMFR: preF=13.9-26.5; postF=9.3-16.0); additional injections did not further elevate antibody levels (Fig 1A and 2A). One mRNA-1653 injection boosted hMPV and PIV3 nAb titers (GMFR over baseline: hMPV-A=2.9-6.1; hMPV-B=6.2-13.2; PIV3=2.8-3.0) and preF and postF bAb concentrations (GMFR: hMPV preF=5.3-6.1; postF=4.6-6.5 and PIV3 preF=13.9-14.2; postF=11.0-12.1); a second injection did not further increase antibody levels (Fig 1B,C and 2B,C). In both trials, bAb responses were generally preF biased. Conclusion In seropositive children aged 12-59 months, mRNA-1345 and mRNA-1653 were well-tolerated and boosted RSV and hMPV plus PIV3 antibodies, respectively, supporting their continued development and that of a combination RSV and hMPV vaccine. Disclosures Matthew D. Snape, MBBS, MD, FRCPCH, FPM, FMedSci, Moderna Biotech UK, Inc.,: Employee|Moderna Biotech UK, Inc.,: Stocks/Bonds Sabine Schnyder Ghamloush, MD, Moderna, Inc.: Employee|Moderna, Inc.: Stocks/Bonds Grace L. Chen, MD, MPH, Moderna, Inc.: Employee|Moderna, Inc.: Stocks/Bonds Rakesh Dhar, MD, Moderna, Inc.: Employee|Moderna, Inc.: Stocks/Bonds Runa Mithani, PharmD, Moderna, Inc: Employee|Moderna, Inc: Stocks/Bonds Vinicius Righi, PharmD, MBA, Moderna, Inc.: Employee|Moderna, Inc.: Stocks/Bonds Louie Morsy, BS, Moderna, Inc.: Employee|Moderna, Inc.: Stocks/Bonds Archana Kapoor, PhD, Moderna, Inc.: Employee|Moderna, Inc.: Stocks/Bonds Bethany Girard, Ph.D., Moderna, Inc.: salary|Moderna, Inc.: Stocks/Bonds Laila El Asmar, PhD, Moderna, Inc.: Employee|Moderna, Inc.: Stocks/Bonds Christine A. Shaw, PhD, Moderna, Inc.: Employee|Moderna, Inc.: Stocks/Bonds

1104. Characteristics and Seasonality of Children Hospitalized with Parainfluenza Virus Infections (PIV), including PIV-4, New Vaccine Surveillance Network, 2016–2020

Abstract Background Human parainfluenza viruses (PIV) are a leading cause of pediatric acute respiratory infection (ARI), yet limited data exist on PIV-associated hospitalizations, particularly from serotype PIV-4, a serotype reported to cause severe disease. We describe the characteristics and seasonality of PIV-associated ARI hospitalizations, focusing on PIV-4. Methods During 12/01/2016-03/31/2020, we enrolled children hospitalized with fever or respiratory symptoms at the 7 children’s hospitals of the CDC’s New Vaccine Surveillance Network. Molecular testing for rhino/enterovirus (RV/EV), respiratory syncytial virus (RSV), influenza virus, PIV (serotypes 1-4), human metapneumovirus, and adenovirus were performed on mid-turbinate nasal or throat specimens collected from enrolled children. Parent interviews and chart abstractions were performed. Results A PIV was detected in 1,004 (6%) of 16,282 enrolled children. A PIV and another respiratory virus (most frequently with RV/EV or RSV) were detected in 336 (33%) children; 668 (67%) children tested positive for only 1 PIV (no co-detection of any other virus or co-detected PIV), including PIV-1 (33%), PIV-2 (15%), PIV-3 (40%), and PIV-4 (12%) (Table 1). Among children with only 1 PIV detected, children with PIV-4 infection were older (mean age 3.1 y) and more likely to have an underlying medical condition than children infected with other PIV serotypes. Discharge diagnoses of asthma, otitis media, and pneumonia were most common in children with PIV-4, and croup most common in those with PIV-1 & 2 (Table 2). PIV-1 had biennial fall seasonality; PIV-2 was detected mostly during the fall of 2018; PIV-3 detections peaked annually in the spring; and PIV-4 was detected year-round (Figure). One in 6 children required ICU care.Figure.Parainfluenza virus (PIV) detections from hospitalized children by month and PIV serotype, New Vaccine Surveillance Network, 2016–2020 Conclusion PIV infections were detected in 6% of ARI hospitalizations and occurred throughout the year, with seasonal variation in serotype dominance. Half of children with PIV-related hospitalizations were healthy (without underlying medical conditions). Severe disease (requirement for ICU care) was common in all PIV-associated hospitalizations. Research on PIV vaccines is warranted, with consideration to include PIV-4. Disclosures John V. Williams, MD, Merck: Grant/Research Support|Quidel: Board Member Marian G. Michaels, MD, MPH, Merck: Grant/Research Support|Viracor: Grant/Research Support Janet A. Englund, MD, Ark Biopharma: Advisor/Consultant|AstraZeneca: Advisor/Consultant|AstraZeneca: Grant/Research Support|GlaxoSmithKline: Grant/Research Support|Meissa Vaccines: Advisor/Consultant|Merck: Grant/Research Support|Moderna: Advisor/Consultant|Moderna: Grant/Research Support|Pfizer: Advisor/Consultant|Pfizer: Grant/Research Support|Sanofi Pasteur: Advisor/Consultant Mary A. Staat, MD, MPH, CDC: Grant/Research Support|Cepheid: Grant/Research Support|Merck: Grant/Research Support|NIH: Grant/Research Support|Pfizer: Grant/Research Support|Up-To-Date: Honoraria Elizabeth P. Schlaudecker, MD, MPH, Pfizer: Grant/Research Support|Sanofi Pasteur: Advisor/Consultant Rangaraj Selvarangan, BVSc, PhD, D(ABMM), FIDSA, FAAM, Abbott: Honoraria|Altona Diagnostics: Grant/Research Support|Baebies Inc: Advisor/Consultant|BioMerieux: Advisor/Consultant|BioMerieux: Grant/Research Support|Bio-Rad: Grant/Research Support|Cepheid: Grant/Research Support|GSK: Advisor/Consultant|Hologic: Grant/Research Support|Lab Simply: Advisor/Consultant|Luminex: Grant/Research Support Natasha B. Halasa, MD, MPH, Merck: Grant/Research Support|Quidell: Grant/Research Support|Quidell: donation of kits|Sanofi: Grant/Research Support|Sanofi: vaccine support Geoffrey A. Weinberg, MD, Merck & Co: Honoraria

1754. Parainfluenza Virus Co-detection With Respiratory Syncytial Virus or Influenza is Associated with Higher Odds of Hospitalization in Children < 2 Years Old, New Vaccine Surveillance Network (NVSN), 2016–2020

Abstract Background Parainfluenza virus (PIV) is an important cause of acute respiratory illness in children; however, scarce data have been reported on the clinical significance of PIV co-detection with other viruses. Methods Between 12/01/2016 and 03/31/2020, we conducted active surveillance for children who presented to the emergency department or were hospitalized with fever or respiratory symptoms at seven U.S. medical centers within NVSN. Demographic and clinical data were collected through parent/guardian interviews and chart abstraction. Nasal and/or throat swabs were tested for PIV types 1–4; respiratory syncytial virus (RSV); rhinovirus or enterovirus (RV/EV); adenovirus (AdV); common cold coronaviruses (ccCoV) 229E, HKU1, NL63, and OC43; SARS-CoV-2; influenza (Flu) A, B, and C; and human metapneumovirus (hMPV). We used a generalized linear mixed-effects model with a logit link to compare the odds of hospitalization between children < 2 years old with PIV-only detection and those with PIV co-detection. Our analysis included age, underlying conditions, and preterm birth as fixed effects and study site as a random effect. Table 1. Comparison of the demographic and clinical characteristics of children with PIV-only detection and those with respiratory virus co-detection, NVSN, 2016–2020. Figure 1. Relative frequencies of respiratory viruses co-detected with PIV in U.S. children <2 years old, stratified by highest* level of care received, NVSN, 2016–2020. Abbreviations: PIV, parainfluenza virus; RV/EV, rhinovirus or enterovirus; RSV, respiratory syncytial virus; AdV, adenovirus; ccCoV, common cold coronaviruses; hMPV, human metapneumovirus; Flu, influenza. Results Of 17,850 children tested for PIV, 1,641 (9.2%) were positive: 1,116 (68.0%) with PIV-only detection and 525 (32.0%) with PIV co-detected with at least one other virus. The demographic and clinical characteristics of children with single PIV detection and those with co-detection are compared in Table 1. Notably, children with PIV co-detection were more likely to attend daycare, preschool, or school. Compared with PIV-only detection, the odds of hospitalization were higher for PIV/RSV (OR=2.18, 95% CI: 1.28–3.76, p=.004) and PIV/Flu (OR=5.61, 95% CI: 1.55–26.60, p=0.014), lower for PIV/AdV(OR=0.48, 95% CI: 0.25–0.89, p=.024), and comparable for the remaining pairs including PIV/RV/EV, the most common co-detection(OR=1.18, 95% CI: 0.86–1.58, p=0.27; Figure 1; Table 2). Table 2. Multivariable logistic regression model of hospitalization in U.S. children with PIV-only detection and those with PIV co-detection with one other respiratory virus, NVSN, 2016–2020. Conclusion The association between PIV co-detection and odds of hospitalization was virus-specific for children < 2 years old. Further investigation is warranted to determine if these findings are driven by RSV and influenza's impact on illness severity or if they are indicative of a potential virus–virus interaction that exacerbates PIV disease. Disclosures Janet A. Englund, MD, Ark Biopharma: Advisor/Consultant|AstraZeneca: Advisor/Consultant|AstraZeneca: Grant/Research Support|GlaxoSmithKline: Grant/Research Support|Meissa Vaccines: Advisor/Consultant|Merck: Grant/Research Support|Moderna: Advisor/Consultant|Moderna: Grant/Research Support|Pfizer: Advisor/Consultant|Pfizer: Grant/Research Support|Sanofi Pasteur: Advisor/Consultant John V. Williams, MD, Merck: Grant/Research Support|Quidel: Board Member Marian G. Michaels, MD, MPH, Merck: Grant/Research Support|Viracor: Grant/Research Support Elizabeth P. Schlaudecker, MD, MPH, Pfizer: Grant/Research Support|Sanofi Pasteur: Advisor/Consultant Mary A. Staat, MD, MPH, CDC: Grant/Research Support|Cepheid: Grant/Research Support|Merck: Grant/Research Support|NIH: Grant/Research Support|Pfizer: Grant/Research Support|Up-To-Date: Honoraria Pedro A. Piedra, MD, Ark Bioscience: Advisor/Consultant|Ark Bioscience: Grant/Research Support|GSK: Grant/Research Support|Icosavax: Advisor/Consultant|Icosavax: Grant/Research Support|Mapp Biologics: Grant/Research Support|Meissa Vaccines: Grant/Research Support|Moderna: Advisor/Consultant|Novavax: Advisor/Consultant|Novavax: Grant/Research Support|Sanofi-Pasteur: Grant/Research Support|Shionogi: Advisor/Consultant|Shionogi: Grant/Research Support|Takeda: Advisor/Consultant Rangaraj Selvarangan, BVSc, PhD, D(ABMM), FIDSA, FAAM, Abbott: Honoraria|Altona Diagnostics: Grant/Research Support|Baebies Inc: Advisor/Consultant|BioMerieux: Advisor/Consultant|BioMerieux: Grant/Research Support|Bio-Rad: Grant/Research Support|Cepheid: Grant/Research Support|GSK: Advisor/Consultant|Hologic: Grant/Research Support|Lab Simply: Advisor/Consultant|Luminex: Grant/Research Support Natasha B. Halasa, MD, MPH, Merck: Grant/Research Support|Quidell: Grant/Research Support|Quidell: donation of kits|Sanofi: Grant/Research Support|Sanofi: vaccine support Geoffrey A. Weinberg, MD, Merck & Co: Honoraria

Positive and negative viral associations in patients with acute respiratory tract infections in primary care: the ECOVIR study.

Acute respiratory infections (ARIs) are the most common viral infections encountered in primary care settings. The identification of causal viruses is still not available in routine practice. Although new strategies of prevention are being identified, knowledge of the relationships between respiratory viruses remains limited. ECOVIR was a multicentric prospective study in primary care, which took place during two pre-pandemic seasons (2018-2019 and 2019-2020). Patients presenting to their General practitioner (GP) with ARIs were included, without selecting for age or clinical conditions. Viruses were detected on nasal swab samples using a multiplex Polymerase Chain Reaction test focused on 17 viruses [Respiratory Syncytial Virus-A (RSV-A), RSV-B, Rhinovirus/Enterovirus (HRV), human Metapneumovirus (hMPV), Adenovirus (ADV), Coronaviruses (CoV) HKU1, NL63, 229E, OC43, Influenza virus (H1 and H3 subtypes), Influenza virus B, Para-Influenza viruses (PIVs) 1-4, and Bocavirus (BoV)]. Among the 668 analyzed samples, 66% were positive for at least one virus, of which 7.9% were viral codetections. The viral detection was negatively associated with the age of patients. BoV, ADV, and HRV occurred more significantly in younger patients than the other viruses (p < 0.05). Codetections were significantly associated with RSV, HRV, BoV, hMPV, and ADV and not associated with influenza viruses, CoV, and PIVs. HRV and influenza viruses were negatively associated with all the viruses. Conversely, a positive association was found between ADV and BoV and between PIVs and BoV. Our study provides additional information on the relationships between respiratory viruses, which remains limited in primary care.

Isolation and Identification of a Genotype C Bovine Parainfluenza Virus Type 3 and Its Pathogenicity in Albino Guinea Pigs

Bovine parainfluenza virus type 3 (BPIV3) is a significant pathogen in cattle, causing a range of respiratory symptoms from mild to severe. In this study, we isolated a BPIV3 (BPIV3/SX/2021) strain using Madin-Darby bovine kidney cells from a lung tissue sample obtained in Shaanxi province. The full length of the BPIV3/SX/2021 strain is 15,474 nt, encoding six structural and three accessory proteins. A phylogenetic analysis revealed that SX2021strain belongs to BPIV3 genotype C, and the isolate shares the highest sequence homology with the other genotype C strains isolated in China. A genetic comparison of the BPIV3c strain isolated in China showed that some amino acid residues have mutations in the F and HN genes that affect the protein structure. Animal challenge experiments showed that experimental infection of guinea pigs with the BPIV3c strain SX2021 displayed depression and dyspnea clinical signs. Histopathological analysis of the lung and trachea showed that morphological changes consisted of lesions of bronchial epithelial cell shedding, alveoli septa thickening, focal fiber hyperplasia, and macrophage infiltration. The virus load analysis indicated that the SX2021 strain displayed a higher replication level in the lungs and trachea. These results provide strong evidence that the evolution of BPIV3, and the isolated BPIV3 strain SX2021 provide useful data to determine the pathogenesis of the virus.

Respiratory Syncytial Virus and Parainfluenza Virus: Characterizing Distinct Clinical Profiles and How They Shift with Bacterial Coinfection

Abstract Objective The objectives are to characterize the distinct clinical profiles of respiratory syncytial virus (RSV) and parainfluenza virus (PIV) infections and how these profiles shift with the addition of bacterial coinfections. Methods This retrospective study analyzed data from more than 1,000 hospitalized children to compare RSV monoinfection with PIV monoinfection, as well as RSV/PIV coinfection with bacteria with monoinfection. Results Significant differences in age distribution (p &lt; 0.001), clinical presentation (p &lt; 0.001), the proportion of pneumonia (p &lt; 0.001), mechanical ventilation (p = 0.004), pediatric intensive care unit (PICU) admission (p = 0.001), and duration of hospitalization (p = 0.003) were observed between RSV and PIV monoinfections. Children with bacterial coinfections had a higher proportion of cough (p = 0.004), wheezing (p = 0.003), rales (p = 0.002), pneumonia (p = 0.002), and PICU admission (p = 0.021) than PIV monoinfection. Notably, the duration of hospitalization for children with bacterial coinfections was longer than that of those infected with a single PIV infection, with a statistically significant difference (p = 0.028). Conclusion Compared with PIV, RSV was more likely to cause severe respiratory tract infections. Coinfection of PIV with bacteria may have exacerbated the severity of acute respiratory tract infections and worsened the symptoms.

Circulation of Parainfluenza Virus in Nashville, Tennessee Before and During the COVID-19 Pandemic

Abstract Background During the COVID-19 pandemic, nonpharmaceutical interventions (NPIs) were introduced and altered circulation of common respiratory viruses, including the parainfluenza viruses (PIVs). Our aim was to describe the circulation of PIVs in Nashville, Tennessee before and during the pandemic. Methods We retrieved records of all respiratory specimens tested using a clinician-ordered respiratory pathogen panel (RPP) test between 01/01/2018, and 12/01/2022, at Vanderbilt University Medical Center. Using the results of RPP testing, we evaluated the proportion of PIV-positive tests before and during the COVID-19 pandemic, including by age groups (0–4, 5–17, 18–64, ≥65), and by PIV types 1-4. Any individual with a medical encounter &amp;gt;14 days was considered a distinct case. We used 04/01/2020, the day Tennessee’s stay-at-home order took effect, as the start date of the pandemic period. Results Of the 58,001 tests ordered during the 60-month study period, 2,601 (4.5%) tests were positive for PIV. PIV-3 was the most common serotype (1,493, 57.4%), followed by PIV-4 (374,14.4%), PIV-2 (356,13.7%), and PIV-1 (351,13.5%). The median age of those who tested PIV-positive was 2.6 years (IQR, 0.9–7.3 years) and 1,395 (53.6%) were male. PIV-1 and PIV-2 were more common children compared to adults. Before the pandemic, PIV-1 and PIV-2 peaked biennially, with PIV-1 peaking in the fall of odd months and PIV-2 peaking in the fall of even years. (Figure 1). PIV-3 and PIV-4 circulated annually, with PIV-3 peaking in the spring and PIV-4 peaking in the fall. During the pandemic, all PIV serotypes skipped a season, but resurfaced the following season. The distribution of detections across age groups was comparable before and during the pandemic. Figure 1. Monthly proportions of RPP tests positive for parainfluenza virus (PIV) in Nashville, Tennessee before and during the COVID-19 pandemic (January 2018–December 2022). The dashed line represents the first day Tennessee’s stay-at-home order was in effect. Co-detection refers to detection of more than one PIV serotype simultaneously. Conclusion The circulation of PIV in Nashville, Tennessee was disrupted during the COVID-19 pandemic, with PIVs not circulating for an entire season. However, circulation resumed the following season. Further investigation is needed to determine if clinical outcomes differed before and during the pandemic and the influence of NPIs shaped PIV transmission.

Prevalence of respiratory viruses among hospitalized children with lower respiratory tract infections during the COVID-19 pandemic in Wuhan, China

ObjectivesWe aimed to investigate the continuous changes in respiratory virus epidemics in hospitalized children with lower respiratory tract infections (LRTIs) persisting from January 2019 to December 2022 in Wuhan, China. MethodsWe retrospectively enrolled children with LRTIs admitted to the Wuhan Children's Hospital. Specimens were nasopharyngeal aspirates which had been collected and detected the following microorganisms with direct immunofluorescence: influenza virus types A and B, respiratory syncytial virus, parainfluenza virus types 1-3, and adenovirus. We also analyzed demographic data and laboratory test results. ResultsA total of 22,660 patients were enrolled. The total virus detection rate in 2019, 2021, and 2022 significantly declined gradually (36.96% vs 29.47% vs 22.62%, P value < 0.001). All the detected viruses did not follow previously observed seasonal patterns during the COVID-19 pandemic. Children hospitalized for LRTIs were older during the COVID-19 pandemic in contrast to the pre-period, particularly notable in cases attributed to respiratory syncytial virus and parainfluenza virus type 3 infections. ConclusionsThis work adds to our knowledge of the epidemiology characteristics of respiratory viruses spanning the COVID-19 pandemic among children with LRTIs. The circulation of respiratory viruses changed consistently, and active LRTI surveillance in children remains critical for defining the healthcare burden of respiratory viruses.

Vosoritide therapy in children with achondroplasia aged 3−59 months: a multinational, randomised, double-blind, placebo-controlled, phase 2 trial

Vosoritide is a recombinant C-type natriuretic peptide analogue that increases annualised growth velocity in children with achondroplasia aged 5-18 years. We aimed to assess the safety and efficacy of vosoritide in infants and children younger than 5 years. This double-blind, randomised, placebo-controlled, phase 2 trial was done in 16 hospitals across Australia, Japan, the UK, and the USA. Children younger than 60 months with a clinical diagnosis of achondroplasia confirmed by genetic testing and who had completed a baseline growth study or observation period were enrolled into one of three sequential cohorts based on age at screening: 24-59 months (cohort 1); 6-23 months (cohort 2); and 0-5 months (cohort 3). Each cohort included sentinels who received vosoritide to determine appropriate daily drug dose, with the remainder randomly assigned (1:1) within each age stratum (except in Japan, where participants were randomly assigned within each cohort) to receive daily subcutaneous injections of vosoritide (30·0 μg/kg for infants aged 0-23 months; 15·0 μg/kg for children aged 24-59 months) or placebo for 52 weeks. Participants, caregivers, investigators, and the sponsor were masked to treatment assignment. The first primary outcome was safety and tolerability, assessed in all participants who received at least one study dose. The second primary outcome was change in height Z score at 52 weeks from baseline, analysed in all randomly assigned participants. This trial is registered with EudraCT, 2016-003826-18, and ClinicalTrials.gov, NCT03583697. Between May 13, 2018, and March 1, 2021, 75 participants were recruited (37 [49%] females). 11 were assigned as sentinels, whereas 32 were randomly assigned to receive vosoritide and 32 placebo. Two participants discontinued treatment and the study: one in the vosoritide group (death) and one in the placebo group (withdrawal). Adverse events occurred in all 75 (100%) participants (annual rate 204·5 adverse events per patient in the vosoritide group and 73·6 per patient in the placebo group), most of which were transient injection-site reactions and injection-site erythema. Serious adverse events occurred in three (7%) participants in the vosoritide group (decreased oxygen saturation, respiratory syncytial virus bronchiolitis and sudden infant death syndrome, and pneumonia) and six (19%) participants in the placebo group (petit mal epilepsy, autism, gastroenteritis, vomiting and parainfluenza virus infection, respiratory distress, and skull fracture and otitis media). The least-squares mean difference for change from baseline in height Z score between the vosoritide and placebo groups was 0·25 (95% CI -0·02 to 0·53). Children with achondroplasia aged 3-59 months receiving vosoritide for 52 weeks had a mild adverse event profile and gain in the change in height Z score from baseline. BioMarin Pharmaceutical.

Novel high-coverage primers for detection of canine morbillivirus by end-point and real-time RT-PCR assays

Canine distemper virus (CDV) is a major threat to domestic dogs and wildlife worldwide. Molecular assays are the most sensitive and specific tests to diagnose the disease, however, the high CDV genetic variability may compromise laboratory diagnosis. Herein, we designed a high-coverage primer set for end-point (RT-PCR) and real-time (RT-qPCR) for CDV detection. Initially, we collected 194 complete/near-complete CDV genomes (GenBank) and analyzed them for highly conserved regions for primer design. We then assessed the in silico coverage, analytical sensitivity, specificity and diagnostic performance of RT-PCR/RT-qPCR reactions based on our primers. Furthermore, the coverage of our primers, as well as their analytical sensitivity and diagnostic performance, were compared to a commonly used primer set for CDV detection (named PP-I). Our forward (F) and reverse (R) primers fully matched 100 % (194/194) and 99 % (192/194) of the analyzed sequences, whereas the PP-I F and R primers fully matched 15 % (29/194) and 9 % (18/194) sequences, respectively. The detection limit of our RT-PCR and RT-qPCR was equivalent to that of PP-I primers (0.001 TCID50/mL). Out of 70 clinical samples tested, 38 were positive by our RT-PCR/RT-qPCR assays, whereas reactions with primers PP-I failed to detect 9/28 (32 %) positive samples selected for comparison purposes. In addition, our assays did not amplify other canine viruses associated with respiratory and neurological diseases: canine adenovirus 2, canine parainfluenza virus 2, canine herpesvirus 1 and rabies virus. Overall, we describe a high-coverage primer set for CDV detection, which represents an attractive tool for laboratory diagnosis of canine distemper.

Predominance of Canine Parainfluenza Virus and Mycoplasma in Canine Infectious Respiratory Disease Complex in Dogs.

Canine infectious respiratory disease complex (CIRDC) is caused by different viruses and bacteria. Viruses associated with CIRDC include canine adenovirus type 2 (CAV-2), canine distemper virus (CDV), canine influenza virus (CIV), canine herpesvirus type 1 (CHV-1), canine respiratory coronavirus (CRCoV), and canine parainfluenza virus (CPIV). Bacteria associated with CIRDC include Bordetella bronchiseptica, Streptococcus equi subspecies zooepidemicus (S. zooepidemicus), and Mycoplasma spp. The present study examined the prevalence of CIRDC pathogens in specimens received by a Veterinary Diagnostic Laboratory in Georgia, USA., from 2018 to 2022. Out of 459 cases, viral agents were detected in 34% of cases and bacterial agents were detected in 58% of cases. A single pathogen was detected in 31% of cases, while two or more pathogens were identified in 24% of cases. The percentages of viral agents identified were CAV-2 (4%), CDV (3%), CPIV (16%), CRCoV (7%), and CIV (2%). The percentages of bacterial agents were B. bronchiseptica (10%), Mycoplasma canis (24%), Mycoplasma cynos (21%), and S. zooepidemicus (2%). Over the five-year period, the positive cases ranged from 2-4% for CAV-2, 1-7% for CDV, 1-4% for CHV-1, 9-22% for CPIV, 4-13% for CRCoV, and 1-4% for CIV. Overall, the most prevalent pathogens associated with CIRDC were CPIV, M. canis, and M. cynos.

Persistent paramyxovirus infections: in co-infections the parainfluenza virus type 5 persistent phenotype is dominant over the lytic phenotype.

Parainfluenza virus type 5 (PIV5) can either have a persistent or a lytic phenotype in cultured cells. We have previously shown that the phenotype is determined by the phosphorylation status of the phosphoprotein (P). Single amino acid substitutions at critical residues, including a serine-to-phenylalanine substitution at position 157 on P, result in a switch between persistent and lytic phenotypes. Here, using PIV5 vectors expressing either mCherry or GFP with persistent or lytic phenotypes, we show that in co-infections the persistent phenotype is dominant. Thus, in contrast to the cell death observed with cells infected solely with the lytic variant, in co-infected cells persistence is immediately established and both lytic and persistent genotypes persist. Furthermore, 10-20 % of virus released from dually infected cells contains both genotypes, indicating that PIV5 particles can package more than one genome. Co-infected cells continue to maintain both genotypes/phenotypes during cell passage, as do individual colonies of cells derived from a culture of persistently infected cells. A refinement of our model on how the dynamics of virus selection may occur in vivo is presented.

Frequency of respiratory virus-associated infection among children and adolescents from a tertiary-care hospital in Mexico City

Acute respiratory infections (ARIs) are a major cause of morbidity and mortality among children. The causative pathogens show geographic and seasonal variations. We retrospectively evaluated the frequency and seasonality of respiratory pathogens in children and adolescents (age: 0–19 years) with ARIs treated between January 1, 2021, and March 31, 2022, at a single center in Mexico. Out of 2400 patients, 1,603 were diagnosed with SARS-CoV-2 infection and 797 were diagnosed with other common respiratory pathogens (CRPs). Of the 797 patients, 632 were infected with one CRP and 165 with > 2 CRPs. Deaths occurred only in SARS-CoV-2-infected patients. Rhinovirus/Enterovirus, respiratory syncytial virus B, and parainfluenza virus 3 were the most prevalent in cases with single and multiple infections. CRP showed a high frequency between autumn and winter of 2021, with higher incidence of hospitalization compared to COVID-19. The main comorbidities were immunosuppression, cardiovascular disease (CD), and asthma. The frequency of CRPs showed a downward trend throughout the first half of 2021. CRPs increased in single- and co-infection cases between the fourth and fifth waves of COVID-19, probably due to decreased nonpharmaceutical interventions and changes in diagnostic tests. Age, cyanosis (symptom), and immunosuppression (comorbidity) were found to differentiate between SARS-CoV-2 infection and CRP infection.

Development and evaluation of a monoclonal antibody-based blocking ELISA to detect antibodies against the E2 protein of bovine viral diarrhea virus-1

With the rapid development of cattle industry, bovine viral diarrhea virus (BVDV) is becoming widespread in China, which causes serious economic losses to the industry. Effective vaccination and viral surveillance are critical for the prevent and control of BVDV infection. In the present study, the immunogenic domain of E2 protein of BVDV-1 was expressed by prokaryotic pET-28a vector. Monoclonal antibodies (mAbs) against E2 protein were prepared and systemically examined by western blot, immunofluorescence assay, blocking ELISA (bELISA) and virus neutralization test (VNT). The mAb 1E2B3, which showed good reactivity and neutralizing activity to BVDV-1 strains, was selected for ELISA establishment. After a series of screening and optimization, a novel bELISA for highly sensitive and specific detection of BVDV-1 antibodies was established, using HRP-labeled 1E2B3 and recombinant E2 protein. ROC analysis of 91 positive and 84 negative reference bovine serum samples yielded the area under the curve (AUC) of 0.9903. A diagnostic specificity of 96.43 % and a sensitivity of 95.6 % were achieved when the cutoff value was set at 24.31 %. There was no cross reaction to the positive sera of classical swine fever virus (CSFV), BVDV-2, border disease virus (BDV), bovine parainfluenza virus type 3 (BPIV3), infectious bovine rhinotracheitis virus (IBRV), foot-and-mouth disease virus (FMDV), Mycoplasma bovis (M.bovis) and Brucella. The total agreement rate of bELISA with VNT was 93.96 % (249/265). In addition, the result of bELISA was positively correlated with neutralizing antibody titer, and the bELISA could well distinguish the serum samples before and after BVDV vaccination. These results indicate that the established bELISA in this study is specific, sensitive, simple and convenient, which provides technical support for the vaccine efficacy evaluation, prevention and control of BVD in the future.

Validation of a real-time PCR panel for detection and quantification of nine pathogens commonly associated with canine infectious respiratory disease

Canine infectious respiratory disease (CIRD) is a complicated respiratory syndrome in dogs [1–3]. A panel PCR was developed [4] to detect nine pathogens commonly associated with CIRD: Mycoplasma cynos, Mycoplasma canis, Bordetella bronchiseptica; canine adenovirus type 2, canine herpesvirus 1, canine parainfluenza virus, canine distemper virus, canine influenza virus and canine respiratory coronavirus [5–16]. To evaluate diagnostic performance of the assay, 740 nasal swab and lung tissue samples were collected and tested with the new assay, and compared to an older version of the assay detecting the same pathogens except that it does not differentiate the two Mycoplasma species. Results indicated that the new assay had the same level of specificity, but with higher diagnostic sensitivity and had identified additional samples with potential co-infections. To confirm the new assay is detecting the correct pathogens, samples with discrepant results between the two assays were sequence-confirmed. Spiking a high concertation target to samples carrying lower concentrations of other targets was carried out and the results demonstrated that there was no apparent interference among targets in the same PCR reaction. Another spike-in experiment was used to determine detection sensitivity between nasal swab and lung tissue samples, and similar results were obtained.•A nine-pathogen CIRD PCR panel assay had identified 139 positives from 740 clinical samples with 60 co-infections;•High-concentration target does not have apparent effect on detecting low-concentration targets;•Detection sensitivity were similar between nasal swab and lung tissue samples.

Immunogenicity and protective efficacy of a recombinant adenovirus containing the fusion protein of bovine parainfluenza virus type 3 genotype C in mice

Bovine parainfluenza virus type 3 (BPIV3) is a viral respiratory pathogen of cattle that causes substantial economic losses. A replicating-defective recombinant human adenovirus type 5 (HAd5), carrying a fusion protein of BPIV3 genotype C (HAd5-F), was constructed and evaluated for its immunogenicity and protective efficacy in mice. After intramuscular injection with the HAd5-F, the IgG titers against F proteins increased to 1:102,400, and virus-neutralizing titers increased to 1:256, significantly higher than those in the group injected with inactivated BPIV3C in mice (p<0.05). The splenic CD4+/CD8+T lymphocytes and IFN-γ+/IL-4+ cytokine percentages were more significant in the HAd5-F group than those in the control group. A BPIV3C challenge in a mouse model was used to assess protective efficacy of the HAd5-F. The viral loads in the lungs and tracheas of mice immunized with the HAd5-F were significantly lower than those in the control group (p<0.0001). There were no significant histopathological alterations in the lungs of mice vaccinated with the HAd5-F. These findings suggested that the HAd5-F elicited excellent immunity against BPIV3C infection.

Markers of antroponotic viral infections in vervet monkeys arrived from their natural habitat (Tanzania).

Various human viruses have been identified in wild monkeys and in captive primates. Cases of transmission of viruses from wild monkeys to humans and vice versa are known. The aim of this study was to identify markers of anthroponotic viral infections in vervet monkeys (Chlorocebus pygerythrus) arrived from their natural habitat (Tanzania). Fecal samples (n = 56) and blood serum samples (n = 75) obtained from 75 animals, respectively, on days 10 and 23 after admission to the primate center, were tested for the markers of anthroponotic viral infections (Ebola virus, Marburg virus, lymphocytic choriomeningitis, hepatitis C virus, herpes simplex virus (HSV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), parainfluenza types 1 and 3, intestinal adenoviruses, rotaviruses) by enzyme immunoassay (ELISA) and polymerase chain reaction (PCR). Among the examined animals, markers of 6 out of 11 tested viral infections were identified. Detection rates of IgG antibodies to HSV-1,2 (15.9%) and CMV (15.9%) were two times as low as IgG antibodies to EBV (31.8%). Among the markers of respiratory viral infections, IgG antibodies to parainfluenza virus type 1 were found (6.8%). 14.3% of the animals had rotavirus antigen, and 94% had simian adenovirus DNA. Markers of hemorrhagic fevers Ebola, Marburg, LCM, hepatitis C, and type 3 parainfluenza were not detected. When importing monkeys from different regions of the world, an expanded screening for viral infections is needed considering the epidemiological situation both in the country of importation and in the country of destination.