tetano
Editor, Senior Moderator
J Clin Virol. 2015 Aug;69:190-6. doi: 10.1016/j.jcv.2015.06.005. Epub 2015 Jun 10.
[h=1]Pathogen transcriptional profile in nasopharyngeal aspirates of children with acute respiratory tract infection.[/h] Fukutani KF[SUP]1[/SUP], Nascimento-Carvalho CM[SUP]2[/SUP], Van der Gucht W[SUP]3[/SUP], Wollants E[SUP]3[/SUP], Khouri R[SUP]1[/SUP], Dierckx T[SUP]3[/SUP], Van Ranst M[SUP]3[/SUP], Houspie L[SUP]3[/SUP], Bouzas ML[SUP]4[/SUP], Oliveira JR[SUP]4[/SUP], Barral A[SUP]5[/SUP], Van Weyenbergh J[SUP]3[/SUP], de Oliveira CI[SUP]6[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] 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.
[h=4]OBJECTIVES:[/h] Detect viral and bacterial RNA in nasopharyngeal aspirates (NPA) from children aged 6-23 months with ARI using nCounter.
[h=4]STUDY DESIGN:[/h] A custom-designed nCounter probeset containing viral and bacterial targets was tested in NPA of ARI patients.
[h=4]RESULTS:[/h] Initially, spiked control viral RNAs were detectable in ≥6.25ng 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 at RNA level, 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.
[h=4]CONCLUSION:[/h] nCounter is robust and sensitive for the simultaneous detection of viral (both RNA and DNA) and bacterial transcripts in NPA with low RNA input (<10ng). 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.
Copyright ? 2015 Elsevier B.V. All rights reserved.
[h=4]KEYWORDS:[/h] ARI; Diagnostics; RSV; nCounter
PMID: 26209405 [PubMed - in process]
[h=1]Pathogen transcriptional profile in nasopharyngeal aspirates of children with acute respiratory tract infection.[/h] Fukutani KF[SUP]1[/SUP], Nascimento-Carvalho CM[SUP]2[/SUP], Van der Gucht W[SUP]3[/SUP], Wollants E[SUP]3[/SUP], Khouri R[SUP]1[/SUP], Dierckx T[SUP]3[/SUP], Van Ranst M[SUP]3[/SUP], Houspie L[SUP]3[/SUP], Bouzas ML[SUP]4[/SUP], Oliveira JR[SUP]4[/SUP], Barral A[SUP]5[/SUP], Van Weyenbergh J[SUP]3[/SUP], de Oliveira CI[SUP]6[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] 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.
[h=4]OBJECTIVES:[/h] Detect viral and bacterial RNA in nasopharyngeal aspirates (NPA) from children aged 6-23 months with ARI using nCounter.
[h=4]STUDY DESIGN:[/h] A custom-designed nCounter probeset containing viral and bacterial targets was tested in NPA of ARI patients.
[h=4]RESULTS:[/h] Initially, spiked control viral RNAs were detectable in ≥6.25ng 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 at RNA level, 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.
[h=4]CONCLUSION:[/h] nCounter is robust and sensitive for the simultaneous detection of viral (both RNA and DNA) and bacterial transcripts in NPA with low RNA input (<10ng). 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.
Copyright ? 2015 Elsevier B.V. All rights reserved.
[h=4]KEYWORDS:[/h] ARI; Diagnostics; RSV; nCounter
PMID: 26209405 [PubMed - in process]