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. 2019 Sep 3;14(9):e0220908.
doi: 10.1371/journal.pone.0220908. eCollection 2019.

Clinical correlation of influenza and respiratory syncytial virus load measured by digital PCR

Diego R Hijano  1 , Jessica Brazelton de Cardenas  2 , Gabriela Maron  1 , Cherilyn D Garner  2 , Jose A Ferrolino  1 , Ronald H Dallas  1 , Zhengming Gu  2 , Randall T Hayden  2
Affiliations

Clinical correlation of influenza and respiratory syncytial virus load measured by digital PCR

Diego R Hijano et al. PLoS One. .

Abstract

Acute respiratory tract infections are a major cause of respiratory morbidity and mortality in pediatric patients worldwide. However, accurate viral and immunologic markers to predict clinical outcomes of this patient population are still lacking. Droplet digital PCR assays for influenza and respiratory syncytial virus (RSV) were designed and performed in 64 respiratory samples from 23 patients with influenza virus infection and 73 samples from 19 patients with RSV infection. Samples of patients with hematologic malignancies, solid tumors, or sickle cell disease were included. Clinical information from institutional medical records was reviewed to assess disease severity. Samples from patients with fever or respiratory symptoms had a significantly higher viral loads than those from asymptomatic patients. Samples from patients with influenza virus and RSV infection collected at presentation had significantly higher viral loads than those collected from patients after completing a course of oseltamivir or ribavirin, respectively. RSV loads correlated positively with clinical symptoms in patients ≤5 years of age, whereas influenza viral loads were associated with clinical symptoms, irrespective of age. Patients receiving antivirals for influenza and RSV had a significant reduction in viral loads after completing therapy. Digital PCR offers an effective method to monitor the efficacy of antiviral treatment for respiratory tract infections in immunocompromised hosts.

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Conflict of interest statement

Randall T. Hayden has served on the advisory boards for Roche Molecular, Abbott Molecular, and Quidel. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1
Fig 1. Correlation of influenza virus and RSV viral loads with clinical infection.
Viral load and presence of fever, cough, and/or nasal congestion for (A) influenza virus A, (B) influenza virus B, and (C) RSV. Viral load and presence of 1–3 symptoms for patients with (D) influenza A, (E) influenza B, and (F) RSV infection. P values from mixed linear regression model *P < 0.05; **P < 0.01; ***P < 0.001. yo, years old; viral loads (log10 copies/ml). Dotted line represents lower limit of quantification for each assay.
Fig 2
Fig 2. Correlation of influenza viral load with clinical signs/symptoms.
Influenza A viral load and presence of (A) fever, (B) cough, and/or (C) nasal congestion. Influenza B viral load and presence of (D) fever, (E) cough, and/or (F) nasal congestion. RSV viral load and presence of (G) fever, (H) cough, and/or (I) nasal congestion. P values from mixed linear regression model *P < 0.05; **P < 0.01; ***P < 0.001. Viral loads (log10 copies/ml). Dotted line represents lower limit of quantification for each assay.
Fig 3
Fig 3. Correlation of lymphopenia with viral load and clinical symptoms.
Correlation between ALC and (A) influenza A viral load, (C) influenza B viral load, and (E) RSV viral load (E). ALCs and the presence of clinical symptoms for (B) influenza A, (D) influenza B, and (F) RSV. P values from mixed linear regression model *P < 0.05; **P < 0.01; ***P < 0.001. Viral loads (log10 copies/ml).
Fig 4
Fig 4. Influenza and RSV viral loads after antiviral treatment.
Influenza viral load before and after oseltamivir treatment for (A) influenza A and (B) influenza B. (C) RSV viral load before and after ribavirin treatment. *P < 0.05; **P < 0.01; ***P < 0.001. Viral loads (log10 copies/ml); Dotted line represents lower limit of quantification for each assay.
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