scholarly journals 491. Persistence of SARS-CoV-2 Iinfection in Immunocompromised Children

2021 ◽  
Vol 8 (Supplement_1) ◽  
pp. S346-S347
Author(s):  
Susan Dolan ◽  
Jean Mulcahy Levy ◽  
Angela Moss ◽  
Kelly Pearce ◽  
Molly Butler ◽  
...  
Keyword(s):  
Viral Load ◽  
Immunosuppressive Treatment ◽  
Viral Persistence ◽  
Research Support ◽  
Repeat Testing ◽  
Viral Loads ◽  
Negative Results ◽  
Significant Difference ◽  
Over Time ◽  
Research Grant

Abstract Background The temporal dynamics of SARS-CoV-2 infectivity in immunocompromised children (IC) are unknown but may have important infection control implications. We evaluated SARS-CoV-2 viral persistence and assessed factors associated with viral persistence and cycle threshold (CT) values as a surrogate of viral load for IC. Methods We conducted a retrospective cohort study of SARS-CoV-2-positive IC at a large quaternary pediatric hospital from March 2020-2021. Immunocompromised status was defined as primary or secondary/acquired immunodeficiencies due to comorbidities or immunosuppressive treatment. The primary outcome was time to first-of-two consecutively negative SARS-CoV-2 PCR tests ≥ 24 hours apart. Polymerase chain reaction (PCR) testing of sequential patient samples was conducted using the Centers for Disease Control 2019-nCoV Real-Time RT-PCR Diagnostic Panel (CDC assay). Chi-square, Fisher exact, and Wilcoxon tests were used to compare demographic and clinical characteristics. Kaplan-Meier curve median event times and log-rank tests were used to compare outcomes. Subjects without 2 consecutive negative tests censored at the last test. Analyses were conducted using SAS v 9.4. Results Ninety-one children met inclusion criteria, and 67 children had more than 1 test (Figure 1). Median age was 15.5 years (IQR 8-18 yrs), 64% were male, 58% of children were white, and 43% were Latinx. Most (67%) were tested in outpatient settings, and 58% of children were asymptomatic. The median time to two negative tests was 42 days (IQR 25.0,55.0), with no difference in duration of positivity with specific diagnoses, degree of lymphopenia, or symptomatic vs asymptomatic illness. Five of 7 (71%) children with samples available for repeat testing had initial CT values < 30, indicating a moderate to high viral load, and of these, 4 (57%) had repeat testing 21 to 30 days later with CT values < 30 (Figure 2), suggesting persistence of moderate to high viral loads. Figure 1. Plot of immunocompromised children in cohort with positive SARS CoV2 PCR and subsequent testing (n = 67). Timelines of immunocompromised children in cohort with positive SARS CoV2 PCR and subsequent testing, grouped by immunocompromising condition. Each line represents an individual patient. Positive results are shown in light grey, negative results are shown in black. Figure 2. Plot of CT values from SARS-CoV-2 PCR testing over time among children with sequential samples available for retesting (n = 7) Plot of CT values (y axis) from SARS-CoV-2 PCR testing on the CDC assay over time (x axis) in days from initial positive test. Repeated testing which yielded a negative result on the CDC assay or intermittent negative results on clinical testing represented as CT value of 40. Each line represents a unique patient. Conclusion The median duration of viral persistence among IC with SARS-CoV-2 infection was 6 weeks, with no significant difference in immunocompromised diagnoses or clinical presentation, with over half of children with testing on the same platform having moderate to high viral loads after 3 weeks, suggesting potential transmission risk. Disclosures Samuel R. Dominguez, MD, PhD, BioFire Diagnostics (Consultant, Research Grant or Support)DiaSorin Molecular (Consultant)Pfizer (Grant/Research Support) Samuel R. Dominguez, MD, PhD, BioFire (Individual(s) Involved: Self): Consultant, Research Grant or Support; DiaSorin Molecular (Individual(s) Involved: Self): Consultant; Pfizer (Individual(s) Involved: Self): Grant/Research Support Suchitra Rao, MBBS, MSCS, BioFire (Research Grant or Support)

scholarly journals SARS-CoV-2 Persistence in Immunocompromised Children

2021 ◽  
Author(s):  
Susan Dolan ◽  
Jean Mulcahy Levy ◽  
Angla Moss ◽  
Kelly Pearce ◽  
Samuel Dominguez ◽  
...  
Keyword(s):  
Viral Load ◽  
Median Time ◽  
Immunosuppressive Treatment ◽  
Viral Persistence ◽  
High Viral Load ◽  
Viral Loads ◽  
Mild Disease ◽  
Kaplan Meier ◽  
Diagnostic Panel ◽  
Event Times

Introduction/Objectives: We evaluated the length of time immunocompromised children (ICC) remain positive for SARS-CoV-2, identified factors associated with viral persistence and determined cycle threshold (CT) values of children with viral persistence as a surrogate of viral load. Methods: We conducted a retrospective cohort study of ICC at a pediatric hospital from March 2020-2021. Immunocompromised status was defined as primary, secondary or acquired due to medical comorbidities/immunosuppressive treatment. The primary outcome was time to first-of-two consecutive negative SARS-CoV-2 Polymerase chain reaction (PCR) tests at least 24 hours apart. Testing of sequential clinical specimens from the same subject was conducted using the Centers for Disease Control (CDC) 2019-nCoV Real-Time RT-PCR Diagnostic Panel assay. Descriptive statistics, Kaplan-Meier curve median event times and log-rank-sum tests were used to compare outcomes between groups. Results: Ninety-one children met inclusion criteria. Median age was 15.5 years (IQR 8-18 yrs), 64% were male, 58% were white, and 43% were Hispanic/Latinx. Most (67%) were tested in outpatient settings and 58% were asymptomatic. The median time to two negative tests was 42 days (IQR 25.0,55.0), with no differences in median time by illness presentation or level of immunosuppression. Seven children had >1 sample available for repeat testing, and 5/7 (71%) children had initial CT values of <30, (moderate to high viral load); 4 children had CT values of <30 3-4 weeks later, suggesting persistent moderate to high viral loads. Conclusions: Most ICC with SARS-CoV-2 infection had mild disease, with prolonged viral persistence >6 weeks and moderate to high viral load.

scholarly journals 108. Selective Decay of Intact HIV-1 Proviral DNA on Antiretroviral Therapy

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S183-S183
Author(s):  
Rajesh Gandhi ◽  
Joshua Cyktor ◽  
Ronald Bosch ◽  
Hanna Mar ◽  
Gregory Laird ◽  
...  
Keyword(s):  
Antiretroviral Therapy ◽  
Panel Member ◽  
Plasma Viremia ◽  
Research Support ◽  
Review Panel ◽  
Proviral Dna ◽  
Hiv 1 ◽  
Over Time ◽  
Residual Plasma Viremia ◽  
Research Grant

Abstract Background HIV-1 proviruses persist in people on antiretroviral therapy (ART) but most are defective and do not constitute a replication-competent reservoir. The decay of infected cells carrying intact compared with defective HIV-1 proviruses has not been well-defined in people on ART. Methods We separately quantified intact and defective proviruses (using an intact proviral DNA assay), residual plasma viremia, and markers of inflammation and activation in people on long-term ART. Longitudinal measurements were done at three timepoints: timepoint 1 was a median of 7.1 years on ART; timepoint 2 was a median of 3.7 years later; timepoint 3 was a median of 5.5 years after timepoint 1 and a median 12 years after starting ART (Figure 1). Figure 1: Study timepoints Results Among 40 participants tested longitudinally from a median of 7.1 years to 12 years after ART initiation, intact provirus levels declined significantly over time (median half-life 7.1 years; 95% confidence interval [CI], 3.9, 18), whereas defective provirus levels did not decrease. The median half-life of total HIV-1 DNA was 41.6 years (95% CI, 13.6, 75). When we evaluated the change in proviral DNA per year, intact proviral DNA declined significantly more (p&lt; 0.001) than defective proviral DNA (the latter did not change) (Figure 2). The proportion of all proviruses that were intact diminished over time on ART, from about 10% at the first on-ART timepoint to about 5% at the last timepoint (Figure 3). At timepoint 1, intact provirus levels on ART correlated with total HIV-1 DNA and residual plasma viremia, but there was no evidence for associations between intact provirus levels and inflammation or immune activation. Figure 2: Percent change in HIV-1 proviral DNA per year Figure 3: Total HIV-1 proviruses (grey bars) and the percentage of intact proviruses (red lines, displaying median, Q1, Q3) by timepoint. Conclusion Cells containing intact, replication-competent proviruses are selectively lost during suppressive ART. Defining the mechanisms involved should inform strategies to accelerate HIV-1 reservoir depletion. Disclosures Rajesh Gandhi, MD, Merck (Advisor or Review Panel member) Gregory Laird, PhD, Accelevir Diagnostics (Shareholder, Other Financial or Material Support, Employee) Albine Martin, PhD, Accelevir Diagnostics (Shareholder, Other Financial or Material Support, Employee) Bernard Macatangay, MD, Gilead (Grant/Research Support) Joseph J. Eron, MD, Gilead Sciences (Consultant, Research Grant or Support)Janssen (Consultant, Research Grant or Support)Merck (Consultant)ViiV Healthcare (Consultant, Research Grant or Support) Janet Siliciano, PhD, Gilead (Advisor or Review Panel member)US Military HIV Research Program (Advisor or Review Panel member) John Mellors, MD, Abound Bio (Shareholder)Accelevir Diagnostics (Consultant)Co-Crystal Pharmaceuticals (Shareholder)Gilead (Consultant, Grant/Research Support)Merck (Consultant)

scholarly journals 520. Longitudinal Analysis of SARS-CoV-2 Viruses in Hospitalized Adults

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S325-S326
Author(s):  
Lacy Simons ◽  
Ramon Lorenzo-Redondo ◽  
Hannah Nam ◽  
Scott C Roberts ◽  
Michael G Ison ◽  
...  
Keyword(s):  
Viral Load ◽  
Genome Sequencing ◽  
Viral Genome ◽  
Temporal Dynamics ◽  
Population Level ◽  
Hospital Staff ◽  
Therapeutic Interventions ◽  
Viral Loads ◽  
Qrt Pcr ◽  
Over Time

Abstract Background The rapid spread of SARS-CoV-2, the causative agent of Coronavirus disease 2019 (COVID-19), has been accompanied by the emergence of viral mutations, some of which may have distinct virological and clinical consequences. While whole genome sequencing efforts have worked to map this viral diversity at the population level, little is known about how SARS-CoV-2 may diversify within a host over time. This is particularly important for understanding the emergence of viral resistance to therapeutic interventions and immune pressure. The goal of this study was to assess the change in viral load and viral genome sequence within patients over time and determine if these changes correlate with clinical and/or demographic parameters. Methods Hospitalized patients admitted to Northwestern Memorial Hospital with a positive SARS-CoV-2 test were enrolled in a longitudinal study for the serial collection of nasopharyngeal specimens. Swabs were administered to patients by hospital staff every 4 ± 1 days for up to 32 days or until the patients were discharged. RNA was extracted from each specimen and viral loads were calculated by quantitative reverse transcriptase PCR (qRT-PCR). Specimens with qRT-PCR cycle threshold values less than or equal to 30 were subject to whole viral genome sequencing by reverse transcription, multiplex PCR, and deep sequencing. Variant populations sizes were estimated and subject to phylogenetic analysis relative to publicly available SARS-CoV-2 sequences. Sequence and viral load data were subsequently correlated to available demographic and clinical data. Results 60 patients were enrolled from March 26th to June 20th, 2020. We observed an overall decrease in nasopharyngeal viral load over time across all patients. However, the temporal dynamics of viral load differed on a patient-by-patient basis. Several mutations were also observed to have emerged within patients over time. Distribution of SARS-CoV-2 viral loads in serially collected nasopharyngeal swabs in hospitalized adults as determined by qRT-PCR. Samples were collected every 4 ± 1 days (T#1–8) and viral load is displayed by log(copy number). Conclusion These data indicate that SARS-CoV-2 viral loads in the nasopharynx decrease over time and that the virus can accumulate mutations during replication within individual patients. Future studies will examine if some of these mutations may provide fitness advantages in the presence of therapeutic and/or immune selective pressures. Disclosures Michael G. Ison, MD MS, AlloVir (Consultant)

scholarly journals Saliva viral load is a dynamic unifying correlate of COVID-19 severity and mortality

2021 ◽  
Author(s):  
Julio Silva ◽  
Carolina Lucas ◽  
Maria Sundaram ◽  
Benjamin Israelow ◽  
Patrick Wong ◽  
...  
Keyword(s):  
Viral Load ◽  
Disease Severity ◽  
T Cell Subsets ◽  
Temporal Association ◽  
Strongly Correlated ◽  
Immunological Parameters ◽  
Viral Loads ◽  
Patient Demographics ◽  
Over Time

While several clinical and immunological parameters correlate with disease severity and mortality in SARS-CoV-2 infection, work remains in identifying unifying correlates of coronavirus disease 2019 (COVID-19) that can be used to guide clinical practice. Here, we examine saliva and nasopharyngeal (NP) viral load over time and correlate them with patient demographics, and cellular and immune profiling. We found that saliva viral load was significantly higher in those with COVID-19 risk factors; that it correlated with increasing levels of disease severity and showed a superior ability over nasopharyngeal viral load as a predictor of mortality over time (AUC=0.90). A comprehensive analysis of immune factors and cell subsets revealed strong predictors of high and low saliva viral load, which were associated with increased disease severity or better overall outcomes, respectively. Saliva viral load was positively associated with many known COVID-19 inflammatory markers such as IL-6, IL-18, IL-10, and CXCL10, as well as type 1 immune response cytokines. Higher saliva viral loads strongly correlated with the progressive depletion of platelets, lymphocytes, and effector T cell subsets including circulating follicular CD4 T cells (cTfh). Anti-spike (S) and anti-receptor binding domain (RBD) IgG levels were negatively correlated with saliva viral load showing a strong temporal association that could help distinguish severity and mortality in COVID-19. Finally, patients with fatal COVID-19 exhibited higher viral loads, which correlated with the depletion of cTfh cells, and lower production of anti-RBD and anti-S IgG levels. Together these results demonstrated that viral load – as measured by saliva but not nasopharyngeal — is a dynamic unifying correlate of disease presentation, severity, and mortality over time.

scholarly journals 986. Evaluation of Moderate-to-Severe Influenza Disease in Children 6 Months to 8 Years of Age in Colorado

2018 ◽  
Vol 5 (suppl_1) ◽  
pp. S291-S292
Author(s):  
Suchitra Rao ◽  
Molly Lamb ◽  
Angela Moss ◽  
Emad Yanni ◽  
Rafik Bekkat-Berkani ◽  
...  
Keyword(s):  
Risk Difference ◽  
Antimicrobial Use ◽  
Clinical Endpoint ◽  
Research Support ◽  
Mild Disease ◽  
Significant Difference ◽  
Ed Visits ◽  
Emergency Room Care ◽  
Negative Controls ◽  
Research Grant

Abstract Background A clinical endpoint of moderate-to-severe (M/S) influenza has been proposed in children, defined as fever &gt;39°C, otitis media, lower respiratory tract infection, or serious extrapulmonary manifestations. This definition has not been evaluated against clinically relevant outcomes like hospitalization, emergency room care, antimicrobial use, and child/parental absenteeism. Methods We conducted a prospective observational study of children aged 6 months–8 years with influenza at the Children’s Hospital Colorado Emergency Department (ED) and its affiliates during two influenza seasons (2016–2017 and 2017–2018). Children with influenza-like-illness (ILI) were enrolled and tested for influenza by polymerase chain reaction (PCR). Parents of influenza cases and matched influenza-negative controls were contacted 2 weeks later for follow-up. The primary outcome was hospitalization for M/S influenza vs. mild influenza. Secondary outcomes included recurrent ED visits, antimicrobial use, child/parental absenteeism. Interim analyses were conducted using SAS v9.4. Results Among the 1,480 enrolled children with ILI, 410 (28%) tested positive for influenza by PCR. The median age of influenza cases was 4.0 years (IQR 2.2–6.1), and 20% were considered high-risk for influenza complications. Of influenza cases, 284 (69%) met the definition for M/S influenza. Among M/S influenza subjects, 8.4% were hospitalized, compared with 1.6% with mild influenza (risk difference (RD) 6.9%; 95% CI: 3.0–10.8, P &lt; 0.01). Subjects with M/S influenza were more likely to receive antibiotics (RD 12.0%, 95% CI: 3.4–20.6, P &lt; 0.01) with a trend to higher antiviral use (RD 6.9%, 95% CI: −0.7–14.5, P = 0.09). There was no significant difference for recurrent ED visits nor child/parental absenteeism. After adjusting for comorbidities, age, and influenza strain, the relative risk (RR) of hospitalization or recurrent ED visits was higher among those with M/S influenza vs. mild influenza (RR 2.18, 95% CI: 1.02–4.64, P = 0.04). Conclusion Children with M/S influenza have a higher risk of hospitalization compared with mild disease. This proposed definition is a useful clinical endpoint to study the public health and clinical impact of influenza interventions in children. Disclosures S. Rao, GSK: Investigator, Research grant. E. Yanni, GSK: Employee, Salary. R. Bekkat-Berkani, GSK: Employee, Salary. A. Schuind, GSK: Employee, Salary. B. Innis, GSK: Employee, Salary. R. Mistry, GSK: Investigator, Research support. E. J. Asturias, GSK: Investigator, Research grant and Research support.

scholarly journals 580. Hesitancy in Uptake and Recommendation of COVID-19 Vaccines by US Healthcare Workers

2021 ◽  
Vol 8 (Supplement_1) ◽  
pp. S392-S392
Author(s):  
Steven S Spires ◽  
Rebecca Rayburn-Reeves ◽  
Elizabeth Dodds Ashley ◽  
Jenna Clark ◽  
Avani P Desai ◽  
...  
Keyword(s):  
Medical Information ◽  
Vaccine Development ◽  
Vaccine Safety ◽  
Vaccine Hesitancy ◽  
Effective Vaccine ◽  
Research Support ◽  
Hospital Employees ◽  
Significant Difference ◽  
Support Research ◽  
Research Grant

Abstract Background The COVID-19 pandemic has brought vaccination to the forefront of discourse on public health. The rapid speed of COVID-19 vaccine development, utilization of novel technology, and an atmosphere of politicized misinformation have created a perfect storm for vaccine hesitancy. As early adopters of vaccination, HCWs set an example for the general population; as trusted sources of medical information, they educate and inform. However, comparatively little work has investigated HCWs' attitudes toward vaccination and how those attitudes drive their recommendation behavior. Methods We surveyed hospital employees about their personal reasons for hesitancy and beliefs about patient hesitancies and randomly assigned them to see one of three messages aimed at increasing vaccine confidence. Message themes included an appeal to return to normal life (Normalcy), a risk comparison between vaccinating or not (SDT), and an explanation of the speed of safe and effective vaccine development (Process). Results Of the 674 NC hospital employees who completed our survey in February 2021, 98% had been offered the COVID-19 vaccine, and 80% had already accepted. For the 20% who had not received the vaccine, the top reasons for hesitancy involved the speed of development and testing, and concerns of vaccine safety and effectiveness. We also found differences in susceptibility to misinformation and vaccine hesitancy across political affiliation, which was higher in Republicans compared to Democrats. HCWs were generally very comfortable recommending the COVID-19 vaccine to patients and supported the idea of sharing the message they read. Although the risk comparison message was most trusted personally, the process message was rated as both the most helpful to patients and the most likely to be shared with them (see Figure 1). This suggests that what is most appealing on a personal level is not necessarily what a HCW would recommend to their patients. Rating of personal opinions of the passages. On a scale from 1 to 7 with 1 = Strongly Disagree and 7 = Strongly Agree. This chart shows the average message ratings across the board when answering whether they thought the passages were understandable, helpful, correct, believable, and trustworthy. (Error bars are 95% CI) There was no significant difference across the messages. The Process message is seen as most helpful and is most likely to be shared with patient than the other messages On left, the average answer on a scale from 1 to 5 for “Do you think the passage you just read would help your patients feel more comfortable about getting the vaccine?” and on right, the average answer for “Would you share this passage with your patients?” Conclusion HCWs' high uptake and minimal hesitancy in recommending the COVID-19 vaccine is encouraging and merits further exploration for how to increase confidence in HCW who are hesitant to discuss and recommend vaccines to patients, as several highlighted the importance of respecting patient autonomy. Disclosures Rebecca Rayburn-Reeves, PhD, Centene Corporation (Grant/Research Support, Research Grant or Support) Jenna Clark, PhD, Centene Corporation (Grant/Research Support, Research Grant or Support) Jan Lindemans, PhD, Centene Corportation (Grant/Research Support, Scientific Research Study Investigator)

scholarly journals Impact of Viral Load Monitoring on Retention and Viral Suppression: A Regression Discontinuity Analysis of South Africa’s National Laboratory Cohort

2020 ◽  
Vol 189 (12) ◽  
pp. 1492-1501
Author(s):  
Alyssa F Harlow ◽  
Jacob Bor ◽  
Alana T Brennan ◽  
Mhairi Maskew ◽  
William MacLeod ◽  
...  
Keyword(s):  
Viral Load ◽  
South African ◽  
Viral Suppression ◽  
Regression Discontinuity ◽  
Risk Difference ◽  
Repeat Testing ◽  
Viral Loads ◽  
Viral Load Monitoring ◽  
Load Monitoring ◽  
To Receive

Abstract South African guidelines recommend repeat viral load testing within 6 months when human immunodeficiency virus (HIV) viral loads exceed 1,000 copies/mL. We assessed whether South African facilities follow viral load monitoring guidelines and whether guidelines improve HIV-related outcomes, using a regression discontinuity design in a national HIV cohort of 174,574 patients (2013–2015). We assessed whether patients with viral loads just above versus just below 1,000 copies/mL were more likely to receive repeat testing in 6 months, and we compared differences in clinic transfers, retention, and viral suppression. The majority (67%) of patients with viral loads of &gt;1,000 copies/mL did not receive repeat testing within 6 months, and these patients were 8.0% (95% confidence interval (CI): 6.2, 9.7) more likely to receive repeat testing compared with ≤1,000 copies/mL. Eligibility for repeat testing (&gt;1,000 copies/mL) was associated with greater 12-month retention (risk difference = 2.9%, 95% CI: 0.6, 5.2) and combined suppression and retention (risk difference = 5.8%, 95% CI: 3.0, 8.6). Patients with viral loads of &gt;1,000 copies/mL who actually received repeat testing were 85.2% more likely to be both retained and virally suppressed at 12 months (95% CI: 35.9, 100.0). Viral load monitoring might improve patient outcomes, but most patients with elevated viral loads do not receive monitoring within recommended timelines.

scholarly journals Evaluation of viral loads in patients with SARS-CoV-2 Delta variant infection: Higher loads do not translate into different testing scenarios

2021 ◽  
Author(s):  
Juan Luis Gomez Marti ◽  
Ashley Mays ◽  
Melissa McCullough ◽  
Alan Wells ◽  
Tung Phan
Keyword(s):  
Viral Load ◽  
Upper Airway ◽  
The Elderly ◽  
Rt Pcr ◽  
Allegheny County ◽  
Viral Loads ◽  
The Third ◽  
Age Related ◽  
Significant Difference ◽  
Fourth Wave

The Delta SARS-CoV-2 variant is very infectious, and it is spreading quickly during this pandemic. In the study, we compared viral loads in surging cases infected with the SARS-CoV-2 Delta variant in the fourth wave of COVID-19 with the three prior waves. The data comprised viral loads from positive cases detected within the UPMC health care system in Allegheny County, Pennsylvania. A total of 2,059 upper airway samples were collected and tested for SARS-CoV-2 positive by RT-PCR during March 2020-September 2021. We did not observe significant difference in viral load difference between the third (December 2020 - January 2021) and fourth (June 2021 - September 2021) waves; however, they had the higher viral load than the first (March 2020 - June 2020) and second waves (June 2020 - August 2020). We did find an age-related effect with the elderly presenting with lower viral loads, which was also seen in the earlier waves. However, the level of viral load in the fourth wave was not sufficient higher to qualitatively change our expected detected rates using various testing modalities.

scholarly journals 685. Comparison of Singleplex qPCR and the Luminex MAGPIX Platform for the Detection of Viral Pathogens

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S396-S396
Author(s):  
Erin Yepsen ◽  
Zaid Haddadin ◽  
Danielle A Rankin ◽  
Rendie McHenry ◽  
Laura S Stewart ◽  
...  
Keyword(s):  
Viral Load ◽  
False Negative ◽  
Respiratory Pathogen ◽  
Influenza B ◽  
Human Bocavirus ◽  
Lower Viral Load ◽  
Research Support ◽  
Ct Value ◽  
Support Research ◽  
Research Grant

Abstract Background Various respiratory molecular assays are available, each with different characteristics and advantages that make them uniquely valuable. The objective of this study was to compare rates of viral detection using singleplex and multiplex platforms in a research setting. Methods A prospective viral surveillance study was conducted in Davidson County, TN. Infants under one year who presented with fever and/or respiratory symptoms were enrolled from the outpatient, emergency department and inpatient settings. Nasal swabs were collected and tested for influenza A (FluA), influenza B (FluB), human metapneumovirus (MPV), respiratory syncytial virus A and B (RSVA and RSVB), human adenovirus (AdV), parainfluenza 1, 2, 3, and 4 (PIV1-4) and SARS-2-CoV by both singleplex qPCR and the Luminex NxTAG Respiratory Pathogen and NxTAG CoV Extended panels. The rhinovirus/enterovirus, human bocavirus, Chlamydophila pneumoniae, Mycoplasma pneumoniae and coronavirus HKU1, NL63, 229E and OC43 results from the Luminex panel were excluded because singleplex qPCR was not performed on those targets. For singleplex qPCR results, cycle threshold (Ct) values were used as a surrogate for viral load, with a higher Ct value indicating a lower viral load. Results A total of 112 nasal specimens were tested by both singleplex qPCR and Luminex, of which 65 were positive for at least one virus by either platform and 56 had a virus detected on both platforms (Figure 1). Seven specimens were positive by singleplex qPCR only and two were positive by Luminex only (Figure 1). The targets positive by singleplex qPCR only included FluB, RSVA, AdV and PIV2 and those positive by Luminex only included FluA H1N1 and RSVB (Figure 2). Specimens that were positive only on the singleplex assay had a higher average Ct value than those that were positive on both assays, indicating a lower viral load (Figure 3). Figure 1 Figure 2 Figure 3 Conclusion The multiplex assay identified 89% of the total viruses detected while singleplex qPCR identified 97% of the total viruses detected. Lower viral loads may contribute to false negative results on the multiplex platforms. Future studies with larger sample sizes are needed in order validate our findings. Disclosures Erin Yepsen, BS, Sanofi Pasteur (Grant/Research Support, Research Grant or Support) Zaid Haddadin, MD, CDC (Grant/Research Support, Research Grant or Support)Quidel Corporation (Grant/Research Support, Research Grant or Support)sanofi pasteur (Grant/Research Support, Research Grant or Support) Danielle A. Rankin, MPH, CIC, Sanofi Pasteur (Grant/Research Support, Research Grant or Support) Natasha B. Halasa, MD, MPH, Genentech (Other Financial or Material Support, I receive an honorarium for lectures - it’s a education grant, supported by genetech)Karius (Consultant)Moderna (Consultant)Quidel (Grant/Research Support, Research Grant or Support)Sanofi (Grant/Research Support, Research Grant or Support)

scholarly journals Molecular Detection and Glycoprotein B (UL55) Genotyping of Cytomegalovirus among Sudanese Renal Transplant Recipients

2019 ◽  
Author(s):  
Hind Haidar Ahmed ◽  
Alfadil Elobeid Omer ◽  
Hisham N. Altyab
Keyword(s):  
Viral Load ◽  
Renal Transplant ◽  
Glycoprotein B ◽  
P Value ◽  
Solid Organ ◽  
Renal Transplant Recipients ◽  
Transplant Recipients ◽  
Viral Loads ◽  
Significant Difference ◽  
Cmv Disease

Abstract Background Cytomegalovirus (CMV) is the most common opportunistic pathogen among solid organ transplant recipients’ especially renal transplants with significant morbidity and mortality. This study was designed to detect CMV DNA and to determine the frequency of different glycoprotein B (UL55) genotypes among Sudanese renal transplant recipients. Methods One hundred and four renal transplant recipients were included in this study. A blood specimen was collected from each recipient. DNA was extracted from plasma using QIAamp DNA mini kit. CMV amplification and quantification (estimation of viral load) was performed using CMV Real – RT Quant kits. Genotyping of Human CMV gB was carried out by nested PCR and sequencing of the highly diverse region of glycoprotein B. Results Cytomegalovirus (CMV) DNA (viremia) was detected in 40/104 (38.5%) of renal transplant recipients. The average of CMV DNA viral load was 358 x104 copies/ml (6.5 log10) ranged from 62 copies/ml (1.8 log 10) to 1.43x108 copies/ml (9 log10). CMV viremia was detected in (60%) of recipients of less than 1-12 months, (17%) of 13-24, (10%) of 25-36, (5%) of 37- 48 and (8%) in more than 48 months post-transplantation with no significant association (P. value = 0.296) between CMV viremia and post renal transplantation time. The association between the type of immunosuppressive drugs and high viral loads (more than 1000 copies /ml) showed a significant difference (P. value =0.05). The correlation between CMV loads of more than 1000 copies/ml and the presence of symptoms of CMV disease were highly significant (P.value =0.00). Fever 7(41%), fever and leucopenia 6(35%) and gastrointestinal disease 4(24%) were the most common presenting symptoms of CMV disease. CMV-genotyping revealed 8 cases (80%) for gB3, and 2 cases (20%) for gB4 genotypes. The most frequent genotype among Sudanese renal transplant recipients was gB3 and no mixed genotypes were observed. Conclusions The frequency of CMV DNA is high among Sudanese renal transplant recipients. CMV viremia viral loads were slightly lower in asymptomatic patients. CMV gB3 is the most predominant glycoprotein B genotype in Sudanese renal transplant recipients.

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