J Clin Virol. 2017 Sep 1;96:12-16. doi: 10.1016/j.jcv.2017.08.014. [Epub ahead of print]
A novel real-time RT-PCR assay for influenza C tested in Peruvian children.
Howard LM1, Johnson M2, Gil AI3, Pekosz A4, Griffin MR5, Edwards KM1, Lanata CF6, Grijalva CG5, Williams JV7; RESPIRA-PERU Group.
Author information
Abstract
BACKGROUND:
Influenza C virus (ICV) is associated with acute respiratory illness. Yet ICV remains under recognized, with most previous studies using only culture to identify cases.
OBJECTIVES:
To develop a sensitive and specific real-time RT-PCR assay for ICV that allows for rapid and accurate detection in a clinical or research setting.
STUDY DESIGN:
Multiple ICV sequences obtained from GenBank were analyzed, including 141 hemagglutinin-esterase (HE), 106 matrix (M), and 97 nucleoprotein (NP) sequences. Primers and probes were designed based on conserved regions. Multiple primer-probe sets were tested against multiple ICV strains.
RESULTS:
The ICV M and NP genes offered the most conserved sequence regions. Primers and probes based on newer sequence data offered enhanced detection of ICV, especially for low titer specimens. An NP-targeted assay yielded the best performance and was capable of detecting 10-100 RNA copies per reaction. The NP assay detected multiple clinical isolates of ICV collected in a field epidemiology study conducted in Peru.
CONCLUSIONS:
We report a new real-time RT-PCR assay for ICV with high sensitivity and specificity.
Copyright ? 2017 Elsevier B.V. All rights reserved.
KEYWORDS:
Diagnostics; Influenza C; Real-time RT-PCR
PMID: 28917132 DOI: 10.1016/j.jcv.2017.08.014
A novel real-time RT-PCR assay for influenza C tested in Peruvian children.
Howard LM1, Johnson M2, Gil AI3, Pekosz A4, Griffin MR5, Edwards KM1, Lanata CF6, Grijalva CG5, Williams JV7; RESPIRA-PERU Group.
Author information
Abstract
BACKGROUND:
Influenza C virus (ICV) is associated with acute respiratory illness. Yet ICV remains under recognized, with most previous studies using only culture to identify cases.
OBJECTIVES:
To develop a sensitive and specific real-time RT-PCR assay for ICV that allows for rapid and accurate detection in a clinical or research setting.
STUDY DESIGN:
Multiple ICV sequences obtained from GenBank were analyzed, including 141 hemagglutinin-esterase (HE), 106 matrix (M), and 97 nucleoprotein (NP) sequences. Primers and probes were designed based on conserved regions. Multiple primer-probe sets were tested against multiple ICV strains.
RESULTS:
The ICV M and NP genes offered the most conserved sequence regions. Primers and probes based on newer sequence data offered enhanced detection of ICV, especially for low titer specimens. An NP-targeted assay yielded the best performance and was capable of detecting 10-100 RNA copies per reaction. The NP assay detected multiple clinical isolates of ICV collected in a field epidemiology study conducted in Peru.
CONCLUSIONS:
We report a new real-time RT-PCR assay for ICV with high sensitivity and specificity.
Copyright ? 2017 Elsevier B.V. All rights reserved.
KEYWORDS:
Diagnostics; Influenza C; Real-time RT-PCR
PMID: 28917132 DOI: 10.1016/j.jcv.2017.08.014