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.
J Clin Virol
; 106: 34-40, 2018 09.
Article
en En
| MEDLINE
| ID: mdl-30031351
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. STUDYDESIGN:
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%CI63.3%-82.9% RSV-A) and 77.6% (95%CI66.3%-85.9% RSV-B); specificity was 98.4% (95%CI96.8%-99.2% RSV-A) and 97.8% (95%CI96.0%-98.8% RSV-B); positive predictive value was 87.3% (95%CI76.9%-93.4% RSV-A) and 82.5% (95%CI71.4%-90.0% RSV-B) and negative predictive value was 96.1% (95%CI94.1%-97.5% RSV-A), and 96.9% (95%CI95.1%-98.2% RSV-B). Accuracy was 95.2% (95%CI93.1%-96.7%) for RSV-A and 95.3% (95%CI93.3%-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.Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Infecciones del Sistema Respiratorio
/
ARN Viral
/
Técnicas de Diagnóstico Molecular
/
Reacción en Cadena de la Polimerasa Multiplex
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
/
Observational_studies
/
Prevalence_studies
/
Prognostic_studies
/
Risk_factors_studies
Límite:
Female
/
Humans
/
Infant
/
Male
País como asunto:
America do sul
/
Brasil
Idioma:
En
Año:
2018
Tipo del documento:
Article