Simultaneous detection of influenza A/B, respiratory syncytial virus, and SARS-CoV-2 in nasopharyngeal swabs by one-tube multiplex reverse transcription polymerase chain reaction

The treatment and outcome of respiratory virus infections differ. SARS-CoV-2, as well as other respiratory viruses such as influenza virus (A and B) and respiratory syncytial virus (RSV), require simultaneous, cost-effective, and rapid differential detection. We used a gold standard fivetarget singl...

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Published in:Tropical medicine and infectious disease 2023-06, Vol.8 (6), p.1-16
Main Authors: Alotaibi, Bader S, Tantry, Bilal Ahmad, Bandy, Altaf, Ahmad, Reyaz, Khursheed, Syed Quibtiya, Ahmad, Arshid, Hakami, Mohammed Ageeli, Shah, Naveed Nazir
Format: Article
Language:English
Subjects:
2020-
5-PLEX assay
COVID-19 Pandemic
Diagnostic use
Epidemics
Genetic research
Genetic transcription
Health aspects
Influenza
influenza A
influenza B
Mortality
Polymerase chain reaction
real-time reverse transcription
Respiratory infections
Respiratory syncytial virus
RSV
SARS-CoV-2
Severe acute respiratory syndrome
Severe acute respiratory syndrome coronavirus 2
Treatment
Viruses
World health
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Summary:The treatment and outcome of respiratory virus infections differ. SARS-CoV-2, as well as other respiratory viruses such as influenza virus (A and B) and respiratory syncytial virus (RSV), require simultaneous, cost-effective, and rapid differential detection. We used a gold standard fivetarget single-step RT-PCR to detect influenza viruses, RSV, and SARS-CoV-2, and this method can be extended to detect influenza virus subtypes. As a result, this five-target single-step RT-PCR method is ideal for differentiating respiratory viruses. The 5' nuclease activity of Taq DNA polymerase is used in the real-time reverse transcription PCR assay. The Taq man fast viral 1-step enzyme is a 4 Master mix and five-target primer probe mix that detects influenza A, influenza B, SARS-CoV- 2 ORF1ab, respiratory syncytial viruses A/B and actin. When compared with TaqMan and Invitrogen superscript III Platinum and the Meril Kit for SARS-CoV-2, the assay demonstrated 100% sensitivity, specificity, and amplification efficiency of 90.1% for target genes. In conclusion, our one-tube multiplex RT-PCR assay offers a rapid and reliable method for the simultaneous detection of influenza A/B, RSV, and SARS-CoV-2 from nasopharyngeal swabs. This assay has the potential to enhance diagnostic capabilities and improve public health responses during respiratory outbreaks, enabling timely interventions and informed decision making.
ISSN:2414-6366
2414-6366
DOI:10.3390/tropicalmed8060326