1. Diagnostic accuracy of digital RNA quantification versus real-time PCR for the detection of respiratory syncytial virus in nasopharyngeal aspirates from children with acute respiratory infection.
- Author
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Bouzas ML, Oliveira JR, Queiroz A, Fukutani KF, Barral A, Rector A, Wollants E, Keyaerts E, Van der Gucht W, Van Ranst M, Beuselinck K, de Oliveira CI, Van Weyenbergh J, and Nascimento-Carvalho CM
- Subjects
- Acute Disease epidemiology, Brazil epidemiology, Cross-Sectional Studies, Female, Humans, Infant, Male, Nasopharynx virology, Prospective Studies, Reproducibility of Results, Respiratory Syncytial Virus, Human genetics, Respiratory Syncytial Virus, Human isolation & purification, Respiratory Tract Infections epidemiology, Respiratory Tract Infections virology, Sensitivity and Specificity, Molecular Diagnostic Techniques methods, Multiplex Polymerase Chain Reaction methods, RNA, Viral analysis, Respiratory Tract Infections diagnosis
- Abstract
Background: Virus-specific molecular assays such as real-time polymerase chain reaction (RT-PCR) are regularly used as the gold standard to diagnose viral respiratory tract infections, but simultaneous detection of multiple different pathogens is often challenging. A multiplex digital method of RNA quantification, nCounter (NanoString Technologies), can overcome this disadvantage and identify, in a single reaction, the presence of different respiratory viruses., Objectives: To evaluate the accuracy of nCounter to identify and quantify RSV-A and RSV-B in nasopharyngeal aspirates (NPA) of children (6-23-months-old) with acute respiratory infection., Study Design: NPA was collected at enrolment in a prospective cross-sectional study conducted in Salvador, Brazil. A quantitative RT-PCR with a subgroup-specific primer and probeset for RSV-A and RSV-B was performed in parallel with a customized nCounter probeset containing viral targets in NPA., Results: Of 559 NPA tested, RSV was detected by RT-PCR in 139 (24.9%), by nCounter in 122 (21.8%) and by any method in 158 (28.3%) cases. Compared to the gold standard of qRT-PCR, sensitivity of nCounter was 74.3% (95%CI:63.3%-82.9% RSV-A) and 77.6% (95%CI:66.3%-85.9% RSV-B); specificity was 98.4% (95%CI:96.8%-99.2% RSV-A) and 97.8% (95%CI:96.0%-98.8% RSV-B); positive predictive value was 87.3% (95%CI:76.9%-93.4% RSV-A) and 82.5% (95%CI:71.4%-90.0% RSV-B) and negative predictive value was 96.1% (95%CI:94.1%-97.5% RSV-A), and 96.9% (95%CI:95.1%-98.2% RSV-B). Accuracy was 95.2% (95%CI:93.1%-96.7%) for RSV-A and 95.3% (95%CI:93.3%-96.9%) for RSV-B, while both methods significantly correlated for RSV-A (r = 0.44, p = 8 × 10
-5 ) and RSV-B (r = 0.73, p = 3 × 10-12 ) quantification., Conclusions: nCounter is highly accurate in detecting RSV-A/B in NPA. Robustness and high-throughput multiplexing indicate its use in large-scale epidemiological studies., (Copyright © 2018 Elsevier B.V. All rights reserved.)- Published
- 2018
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