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J Clin Virol . Design and performance of a real-time RT-PCR assay for detection of influenza C viruses

tetano

Editor, Senior Moderator
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[SUP] 1 [/SUP], William G Davis[SUP] 1 [/SUP], Ji Liu[SUP] 1 [/SUP], Beth K Thielen[SUP] 2 [/SUP], Sarah Bistodeau[SUP] 3 [/SUP], Brian Lynch[SUP] 1 [/SUP], Christine M Warnes[SUP] 1 [/SUP], Jimma Liddell[SUP] 1 [/SUP], Anna K Strain[SUP] 3 [/SUP], Jaime Christensen[SUP] 3 [/SUP], Phili Wong[SUP] 1 [/SUP], Natasha Burnett[SUP] 1 [/SUP], Todd C Davis[SUP] 1 [/SUP], Marie K Kirby[SUP] 4 [/SUP]



Affiliations
Abstract

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.

 
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