Article
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
Registro en:
BOUZAS, Maiara Lanna Souza Bacelar et al. 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. Journal of Clinical Virology, v. 106, p. 34-40, 2018.
1386-6532
10.1016/j.jcv.2018.07.003
Autor
Bouzas, Maiara Lanna Souza Bacelar
Oliveira, Juliana R.
Queiroz, Artur Trancoso Lopo de
Fukutani, Kiyoshi Ferreira
Barral, Aldina Maria Prado
Rector, Annabel
Wollants, Elke
Keyaerts, Els
Van der Gucht, Winke
Van Ranst, Marc
Beuselinck, Kurt
Oliveira, Camila Indiani de
Van Weyenbergh, Johan
Carvalho, Cristiana Maria Costa Nascimento
The Acute Respiratory Infection and Wheeze Study Group Phase I and II
Resumen
Ethics Committee from the Federal University of Bahia (no 067/2009). 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.
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Virology Branch; Universidade de São Paulo (USP); Instituto Adolfo Lutz (IAL); Centers for Disease Control and Prevention; STD/AIDS Reference and Training Centre; Pennsylvania State University; National Institutes of Health; Universidade Estadual Paulista (Unesp) (2009-01-01)Human respiratory syncytial virus (HRSV) is the major cause of lower respiratory tract infections in children under 5 years of age and the elderly, causing annual disease outbreaks during the fall and winter. Multiple ... -
Positive selection results in frequent reversible amino acid replacements in the G protein gene of human respiratory syncytial virus
Virology Branch; Universidade de São Paulo (USP); Instituto Adolfo Lutz (IAL); Centers for Disease Control and Prevention; STD/AIDS Reference and Training Centre; Pennsylvania State University; National Institutes of Health; Universidade Estadual Paulista (Unesp) (2009-01-01)Human respiratory syncytial virus (HRSV) is the major cause of lower respiratory tract infections in children under 5 years of age and the elderly, causing annual disease outbreaks during the fall and winter. Multiple ...