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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Bouzas, Maiara L. [1] ; Oliveira, Juliana R. [1] ; Queiroz, Artur [2] ; Fukutani, Kiyoshi F. [1, 3] ; Barral, Aldina [2, 1, 4] ; Rector, Annabel [5] ; Wollants, Elke [5] ; Keyaertse, Els [5, 6] ; Van der Gucht, Winke [5] ; Van Ranst, Marc [5, 6] ; Beuselinck, Kurt [6] ; de Oliveira, I, Camila ; Van Weyenbergh, Johan [5] ; Nascimento-Carvalho, Cristiana M. [7, 8] ; Grp, Acute Resp Infect Wheeze Study
Total Authors: 15
[1] Univ Fed Bahia, Sch Med, Postgrad Program Hlth Sci, Salvador, BA - Brazil
[2] I, Fundacao Oswaldo Cruz FIOCRUZ, Ctr Pesquisas Goncalo Moniz CPqGM, Salvador, BA - Brazil
[3] Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Biochem Immunol & Cell Biol, Ribeirao Preto - Brazil
[4] Univ Fed Bahia, Sch Med, Dept Pathol, Salvador, BA - Brazil
[5] Katholieke Univ Leuven, Rega Inst Med Res, Lab Clin & Epidemiol Virol, Dept Microbiol & Immunol, Leuven - Belgium
[6] Univ Hosp Leuven, Dept Lab Med, Leuven - Belgium
[7] de Oliveira, Camila, I, Univ Fed Bahia, Sch Med, Postgrad Program Hlth Sci, Salvador, BA - Brazil
[8] Univ Fed Bahia, Sch Med, Dept Pediat, Salvador, BA - Brazil
Total Affiliations: 8
Document type: Journal article
Source: Journal of Clinical Virology; v. 106, p. 34-40, SEP 2018.
Web of Science Citations: 3

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 x 10(-5)) and RSV-B (r = 0.73, p = 3 x 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. (AU)

FAPESP's process: 17/03491-6 - RIDCs: Research, Innovation and Dissemination Centers
Grantee:Kiyoshi Ferreira Fukutani
Support type: Scholarships in Brazil - Post-Doctorate