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https://www.arca.fiocruz.br/handle/icict/13752
PATHOGEN TRANSCRIPTIONAL PROFILE IN NASOPHARYNGEAL ASPIRATES OF CHILDREN WITH ACUTE RESPIRATORY TRACT INFECTION
Author
Fukutani, Kiyoshi Ferreira
Carvalho, Cristiana Maria Costa Nascimento
Gucht, Winke Van der
Wollants, Elke
Cunha, Antonio Ricardo Khouri
Dierckx, Tim
Van Ranst, Marc
Houspie, Lieselot
Bouzas, Maiara Lanna Souza Bacelar
Oliveira, Juliana R.
Barral, Aldina Maria Prado
Van Weyenbergh, Johan Jozef Rosa Maria
Oliveira, Camila Indiani de
Carvalho, Cristiana Maria Costa Nascimento
Gucht, Winke Van der
Wollants, Elke
Cunha, Antonio Ricardo Khouri
Dierckx, Tim
Van Ranst, Marc
Houspie, Lieselot
Bouzas, Maiara Lanna Souza Bacelar
Oliveira, Juliana R.
Barral, Aldina Maria Prado
Van Weyenbergh, Johan Jozef Rosa Maria
Oliveira, Camila Indiani de
Affilliation
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil.
Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil / Federal University of Bahia. School of Medicine. Department of Pediatrics. Salvador, BA, Brasil.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil.
Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil.
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil / Federal University of Bahia. School of Medicine. Department of Pathology. School of Medicine. Salvador, BA, Brasil / Instituto de Investigação em Imunologia. São Paulo, SP, Brasil.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil / Instituto de Investigação em Imunologia. São Paulo, SP, Brasil.
Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil / Federal University of Bahia. School of Medicine. Department of Pediatrics. Salvador, BA, Brasil.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil.
Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil.
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil / Federal University of Bahia. School of Medicine. Department of Pathology. School of Medicine. Salvador, BA, Brasil / Instituto de Investigação em Imunologia. São Paulo, SP, Brasil.
Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia School of Medicine. Post-Graduation Program in Health Sciences. Salvador, BA, Brasil / Instituto de Investigação em Imunologia. São Paulo, SP, Brasil.
Abstract
Background: Acute respiratory tract infections (ARI) present a significant morbidity and pose a global
health burden. Patients are frequently treated with antibiotics although ARI are most commonly caused
by virus, strengthening the need for improved diagnostic methods.
Objectives: Detect viral and bacterial RNA in nasopharyngeal aspirates (NPA) from children aged 6–23
months with ARI using nCounter.
Study design: A custom-designed nCounter probeset containing viral and bacterial targets was tested in
NPA of ARI patients.
Results: Initially, spiked control viral RNAs were detectable in ≥6.25 ng input RNA, indicating absence of
inhibitors in NPA. nCounter applied to a larger NPA sample (n = 61) enabled the multiplex detection of
different pathogens: RNA viruses Parainfluenza virus (PIV 1–3) and RSV A-B in 21%, Human metapneumovirus
(hMPV) in 5%, Bocavirus (BoV), CoV, Influenza virus (IV) A in 3% and, Rhinovirus (RV) in 2% of
samples, respectively. RSV A-B was confirmed by Real Time PCR (86.2–96.9% agreement). DNA virus (AV)
was detected atRNAlevel, reflecting viral replication, in 10% of samples. Bacterial transcripts from Staphylococcus
aureus, Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis, Mycoplasma
pneumoniae and Chlamydophila pneumoniae were detected in 77, 69, 26, 8, 3 and 2% of samples, respectively.
Conclusion: nCounter is robust and sensitive for the simultaneous detection of viral (both RNA and DNA)
and bacterial transcripts in NPA with low RNA input (<10 ng). This medium-throughput technique will
increase our understanding of ARI pathogenesis and may provide an evidence-based approach for the
targeted and rational use of antibiotics in pediatric ARI.
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