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J Clin Virol
. 2025 Sep 25:181:105874.
doi: 10.1016/j.jcv.2025.105874. Online ahead of print. Design and performance of a real-time RT-PCR assay for detection of influenza C viruses
Bo Shu 1 , William G Davis 1 , Ji Liu 1 , Beth K Thielen 2 , Sarah Bistodeau 3 , Brian Lynch 1 , Christine M Warnes 1 , Jimma Liddell 1 , Anna K Strain 3 , Jaime Christensen 3 , Phili Wong 1 , Natasha Burnett 1 , Todd C Davis 1 , Marie K Kirby 4
Affiliations
Influenza C virus (ICV) usually causes a mild upper respiratory tract infection in children and those infected are frequently co-infected with other respiratory viruses. However, there have only been a few hundred documented cases of ICV infection in humans as of the end of 2024. To better understand the epidemiology and clinical impact of ICVs, we developed an influenza C real-time RT-PCR (InfC rRT-PCR) assay that targets a highly conserved region of the matrix gene segment of ICVs. The analytical sensitivity evaluation demonstrated that the InfC rRT-PCR assay was highly sensitive, as it was able to detect as few as five RNA copies per PCR reaction and had robust reactivity over a range of viral RNAs from historical and recent ICVs. The analytical specificity evaluation confirmed the assay did not cross-react with any influenza A or B viruses tested, including several animal-origin viruses, or other common non-influenza respiratory viruses. The performance evaluation on clinical specimens demonstrated the assay was highly sensitive and specific for the detection of ICVs.
Keywords: Diagnostic assay; Influenza C virus; Real-time RT-PCR.
. 2025 Sep 25:181:105874.
doi: 10.1016/j.jcv.2025.105874. Online ahead of print. Design and performance of a real-time RT-PCR assay for detection of influenza C viruses
Bo Shu 1 , William G Davis 1 , Ji Liu 1 , Beth K Thielen 2 , Sarah Bistodeau 3 , Brian Lynch 1 , Christine M Warnes 1 , Jimma Liddell 1 , Anna K Strain 3 , Jaime Christensen 3 , Phili Wong 1 , Natasha Burnett 1 , Todd C Davis 1 , Marie K Kirby 4
Affiliations
- PMID: 41033150
- DOI: 10.1016/j.jcv.2025.105874
Influenza C virus (ICV) usually causes a mild upper respiratory tract infection in children and those infected are frequently co-infected with other respiratory viruses. However, there have only been a few hundred documented cases of ICV infection in humans as of the end of 2024. To better understand the epidemiology and clinical impact of ICVs, we developed an influenza C real-time RT-PCR (InfC rRT-PCR) assay that targets a highly conserved region of the matrix gene segment of ICVs. The analytical sensitivity evaluation demonstrated that the InfC rRT-PCR assay was highly sensitive, as it was able to detect as few as five RNA copies per PCR reaction and had robust reactivity over a range of viral RNAs from historical and recent ICVs. The analytical specificity evaluation confirmed the assay did not cross-react with any influenza A or B viruses tested, including several animal-origin viruses, or other common non-influenza respiratory viruses. The performance evaluation on clinical specimens demonstrated the assay was highly sensitive and specific for the detection of ICVs.
Keywords: Diagnostic assay; Influenza C virus; Real-time RT-PCR.