tetano
Editor, Senior Moderator
Virus Res
. 2024 May 28:199410.
doi: 10.1016/j.virusres.2024.199410. Online ahead of print. Simultaneous and ultrafast detection of pan-SARS-coronaviruses and influenza A/B viruses by a novel multiplex real-time RT-PCR assay
Changping Xu[SUP] 1 [/SUP], Zhengyang Wang[SUP] 2 [/SUP], Beibei Yu[SUP] 3 [/SUP], Zhenhuang Pan[SUP] 4 [/SUP], Jun Ni[SUP] 5 [/SUP], Yan Feng[SUP] 6 [/SUP], Shiwang Huang[SUP] 7 [/SUP], Maomao Wu[SUP] 8 [/SUP], Jiancang Zhou[SUP] 5 [/SUP], Lei Fang[SUP] 9 [/SUP], Zhiwei Wu[SUP] 10 [/SUP]
Affiliations
Here we report an ultrafast quadruplex RT-qPCR assay with robust diagnostic ability to detect and distinguish pan-SARS-CoVs and influenza A/B viruses within 35 min. This quadruplex RT-qPCR assay comprised of one novel RNA-based internal control targeting human β2-microglobulin (B2M) for process accuracy and three newly-designed primers-probe sets targeting the envelope protein (E) of pan-SARS-CoV, matrix protein (MP) of influenza A virus and non-structural (NS) region of influenza B virus. This quadruplex assay exhibited a sensitivity comparable to its singleplex counterparts and a slightly higher to that of the Centers for Disease Control and Prevention-recommended SARS-CoV-2 and influenza A/B assays. The novel assay showed no false-positive amplifications with other common respiratory viruses, and its 95% limits of detection for pan-SARS-CoV and influenza A/B virus was 4.26-4.52 copies/reaction. Moreover, the assay was reproducible with less than 1% coefficient of variation and adaptable testing different clinical and environmental samples. Our ultrafast quadruplex RT-qPCR assay can serve as an attractive tool for effective differentiation of influenza A/B virus and SARS-CoV-2, but more importantly prognose the reemergence/emergence of SARS and novel coronaviruses or influenza viruses from animal spillover.
Keywords: SARS; SARS-CoV-2; co-surveillance; coinfection; influenza; molecular diagnostics; viral spillover.
. 2024 May 28:199410.
doi: 10.1016/j.virusres.2024.199410. Online ahead of print. Simultaneous and ultrafast detection of pan-SARS-coronaviruses and influenza A/B viruses by a novel multiplex real-time RT-PCR assay
Changping Xu[SUP] 1 [/SUP], Zhengyang Wang[SUP] 2 [/SUP], Beibei Yu[SUP] 3 [/SUP], Zhenhuang Pan[SUP] 4 [/SUP], Jun Ni[SUP] 5 [/SUP], Yan Feng[SUP] 6 [/SUP], Shiwang Huang[SUP] 7 [/SUP], Maomao Wu[SUP] 8 [/SUP], Jiancang Zhou[SUP] 5 [/SUP], Lei Fang[SUP] 9 [/SUP], Zhiwei Wu[SUP] 10 [/SUP]
Affiliations
- PMID: 38815870
- DOI: 10.1016/j.virusres.2024.199410
Here we report an ultrafast quadruplex RT-qPCR assay with robust diagnostic ability to detect and distinguish pan-SARS-CoVs and influenza A/B viruses within 35 min. This quadruplex RT-qPCR assay comprised of one novel RNA-based internal control targeting human β2-microglobulin (B2M) for process accuracy and three newly-designed primers-probe sets targeting the envelope protein (E) of pan-SARS-CoV, matrix protein (MP) of influenza A virus and non-structural (NS) region of influenza B virus. This quadruplex assay exhibited a sensitivity comparable to its singleplex counterparts and a slightly higher to that of the Centers for Disease Control and Prevention-recommended SARS-CoV-2 and influenza A/B assays. The novel assay showed no false-positive amplifications with other common respiratory viruses, and its 95% limits of detection for pan-SARS-CoV and influenza A/B virus was 4.26-4.52 copies/reaction. Moreover, the assay was reproducible with less than 1% coefficient of variation and adaptable testing different clinical and environmental samples. Our ultrafast quadruplex RT-qPCR assay can serve as an attractive tool for effective differentiation of influenza A/B virus and SARS-CoV-2, but more importantly prognose the reemergence/emergence of SARS and novel coronaviruses or influenza viruses from animal spillover.
Keywords: SARS; SARS-CoV-2; co-surveillance; coinfection; influenza; molecular diagnostics; viral spillover.