Please use this identifier to cite or link to this item:
https://www.arca.fiocruz.br/handle/icict/28067
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
Diagnóstico
Vírus Respiratório
Detecção de Vírus
Infecção respiratória viral
Transcriptômica
Diagnostics
Respiratory viruses
Virus detection
Viral respiratory infection
Transcriptomics
Author
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
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
Affilliation
Federal University of Bahia. School of Medicine. Postgraduate Program in Health Sciences. Salvador, BA, Brazil.
Federal University of Bahia. School of Medicine. Postgraduate Program in Health Sciences. Salvador, BA, Brazil.
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil.
Federal University of Bahia. School of Medicine. Postgraduate Program in Health Sciences. Salvador, BA, Brazil / University of São Paulo. Ribeirão Preto Medical School. Department of Biochemistry, Immunology, and Cell Biology. Ribeirão Preto, SP, Brazil.
Federal University of Bahia. School of Medicine. Postgraduate Program in Health Sciences. Salvador, BA, Brazil / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia. School of Medicine. Department of Pathology. Salvador, BA, Brazil.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium / University Hospitals Leuven. Department of Laboratory Medicine. Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium / University Hospitals Leuven. Department of Laboratory Medicine. Leuven, Belgium.
University Hospitals Leuven. Department of Laboratory Medicine. Leuven, Belgium.
Federal University of Bahia. School of Medicine. Postgraduate Program in Health Sciences. Salvador, BA, Brazil / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium.
Federal University of Bahia. School of Medicine. Postgraduate Program in Health Sciences. Salvador, BA, Brazil / Federal University of Bahia. School of Medicine. Department of Pediatrics. Salvador, BA, Brazil.
Federal University of Bahia. School of Medicine. Postgraduate Program in Health Sciences. Salvador, BA, Brazil.
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil.
Federal University of Bahia. School of Medicine. Postgraduate Program in Health Sciences. Salvador, BA, Brazil / University of São Paulo. Ribeirão Preto Medical School. Department of Biochemistry, Immunology, and Cell Biology. Ribeirão Preto, SP, Brazil.
Federal University of Bahia. School of Medicine. Postgraduate Program in Health Sciences. Salvador, BA, Brazil / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia. School of Medicine. Department of Pathology. Salvador, BA, Brazil.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium / University Hospitals Leuven. Department of Laboratory Medicine. Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium / University Hospitals Leuven. Department of Laboratory Medicine. Leuven, Belgium.
University Hospitals Leuven. Department of Laboratory Medicine. Leuven, Belgium.
Federal University of Bahia. School of Medicine. Postgraduate Program in Health Sciences. Salvador, BA, Brazil / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil.
Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory for Clinical and Epidemiological Virology. KU Leuven, Leuven, Belgium.
Federal University of Bahia. School of Medicine. Postgraduate Program in Health Sciences. Salvador, BA, Brazil / Federal University of Bahia. School of Medicine. Department of Pediatrics. Salvador, BA, Brazil.
Abstract
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.
Keywords in Portuguese
RSVDiagnóstico
Vírus Respiratório
Detecção de Vírus
Infecção respiratória viral
Transcriptômica
Keywords
RSVDiagnostics
Respiratory viruses
Virus detection
Viral respiratory infection
Transcriptomics
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