Pathogen transcriptional profile in nasopharyngeal aspirates of children with acute respiratory tract infection
| Autor: | Kiyoshi F. Fukutani, Winke Van der Gucht, Lieselot Houspie, Cristiana M. Nascimento-Carvalho, Juliana R. Oliveira, Johan Van Weyenbergh, Elke Wollants, Aldina Barral, Ricardo Khouri, Tim Dierckx, Maiara L. Bouzas, Marc Van Ranst, Camila I. de Oliveira |
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| Rok vydání: | 2015 |
| Předmět: |
Mycoplasma pneumoniae
viruses Biology medicine.disease_cause Sensitivity and Specificity Article Virus Haemophilus influenzae Microbiology stomatognathic system Human metapneumovirus Nasopharynx Virology Streptococcus pneumoniae medicine Humans Diagnostics Respiratory Tract Infections Phylogeny Bacteria Respiratory tract infections Gene Expression Profiling RSV Infant Bacterial Infections ARI biology.organism_classification respiratory tract diseases RNA Bacterial Infectious Diseases Virus Diseases Chlamydophila pneumoniae nCounter Viruses RNA Viral Rhinovirus Multiplex Polymerase Chain Reaction |
| Zdroj: | Journal of Clinical Virology |
| ISSN: | 1386-6532 |
| Popis: | Highlights • nCounter enables detection of pathogen transcripts in NPA with low RNA input. • nCounter detects, in a single reaction, the presence of multiple pathogens in NPA. • nCounter displayed a good agreement with Real-Time PCR for RSV. 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 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. 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 ( |
| Databáze: | OpenAIRE |
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