Design and assessment of a multiplex real-time PCR method for simultaneous detection and differentiation of COVID-19 and Influenza A/B
Background and Objectives: Viral infections of the respiratory system are a major public problem due to their ease of spread, pandemic potential, and significant rate of death. Diagnosing these infections requires laboratory testing, as clinical symptoms alone are often insufficient. Influenza A, I...
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| Format: | Article |
| Language: | English |
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Tehran University of Medical Sciences
2025-04-01
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| Series: | Iranian Journal of Microbiology |
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| Online Access: | https://ijm.tums.ac.ir/index.php/ijm/article/view/5047 |
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| author | Nafiseh Fotros Reihaneh Bashiri Samira Mohammadi-Yeganeh Mahdi Paryan |
| author_facet | Nafiseh Fotros Reihaneh Bashiri Samira Mohammadi-Yeganeh Mahdi Paryan |
| author_sort | Nafiseh Fotros |
| collection | DOAJ |
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Background and Objectives: Viral infections of the respiratory system are a major public problem due to their ease of spread, pandemic potential, and significant rate of death. Diagnosing these infections requires laboratory testing, as clinical symptoms alone are often insufficient. Influenza A, Influenza B, and COVID-19 are common infections that burden the population, especially during winter. We developed a multiplex real-time PCR method to simultaneously detect Influenza A and B, as well as COVID-19. Compared to existing detection kits, it offers higher accuracy, lower costs, and faster results, making it an efficient diagnostic tool.
Materials and Methods: We designed primer/TaqMan probes for the M2 gene of Influenza A, N gene of SARS-CoV-2, and NS1 gene of Influenza B. Reaction components were optimized and functional parameters were tested using standard samples with known viral copy numbers.
Results: The method’s detection limit is 10 copies for Influenza A and B, and 60 for SARS-CoV-2. Sensitivity and specificity for Influenza A are 88% and 100%, for Influenza B, 95.6% and 100%, and for SARS-CoV-2, 90.4% and 100%.
Conclusion: This multiplex real-time PCR method can accurately detect and distinguish SARS-CoV-2, Influenza B, and Influenza A infections.
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| format | Article |
| id | doaj-art-c7c5e2ac1b7546cbbf71d2b452811f18 |
| institution | DOAJ |
| issn | 2008-3289 2008-4447 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Microbiology |
| spelling | doaj-art-c7c5e2ac1b7546cbbf71d2b452811f182025-08-20T03:03:40ZengTehran University of Medical SciencesIranian Journal of Microbiology2008-32892008-44472025-04-0117210.18502/ijm.v17i2.18381Design and assessment of a multiplex real-time PCR method for simultaneous detection and differentiation of COVID-19 and Influenza A/BNafiseh Fotros0Reihaneh Bashiri1Samira Mohammadi-Yeganeh2Mahdi Paryan3Department of Biotechnology, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, IranDepartment of Biotechnology, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, IranMedical Nanotechnology and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, IranDepartment of Research and Development, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran Background and Objectives: Viral infections of the respiratory system are a major public problem due to their ease of spread, pandemic potential, and significant rate of death. Diagnosing these infections requires laboratory testing, as clinical symptoms alone are often insufficient. Influenza A, Influenza B, and COVID-19 are common infections that burden the population, especially during winter. We developed a multiplex real-time PCR method to simultaneously detect Influenza A and B, as well as COVID-19. Compared to existing detection kits, it offers higher accuracy, lower costs, and faster results, making it an efficient diagnostic tool. Materials and Methods: We designed primer/TaqMan probes for the M2 gene of Influenza A, N gene of SARS-CoV-2, and NS1 gene of Influenza B. Reaction components were optimized and functional parameters were tested using standard samples with known viral copy numbers. Results: The method’s detection limit is 10 copies for Influenza A and B, and 60 for SARS-CoV-2. Sensitivity and specificity for Influenza A are 88% and 100%, for Influenza B, 95.6% and 100%, and for SARS-CoV-2, 90.4% and 100%. Conclusion: This multiplex real-time PCR method can accurately detect and distinguish SARS-CoV-2, Influenza B, and Influenza A infections. https://ijm.tums.ac.ir/index.php/ijm/article/view/5047Multiplex;Real-time polymerase chain reaction;SARS-CoV-2;COVID-19;Influenza B;Influenza A |
| spellingShingle | Nafiseh Fotros Reihaneh Bashiri Samira Mohammadi-Yeganeh Mahdi Paryan Design and assessment of a multiplex real-time PCR method for simultaneous detection and differentiation of COVID-19 and Influenza A/B Iranian Journal of Microbiology Multiplex; Real-time polymerase chain reaction; SARS-CoV-2; COVID-19; Influenza B; Influenza A |
| title | Design and assessment of a multiplex real-time PCR method for simultaneous detection and differentiation of COVID-19 and Influenza A/B |
| title_full | Design and assessment of a multiplex real-time PCR method for simultaneous detection and differentiation of COVID-19 and Influenza A/B |
| title_fullStr | Design and assessment of a multiplex real-time PCR method for simultaneous detection and differentiation of COVID-19 and Influenza A/B |
| title_full_unstemmed | Design and assessment of a multiplex real-time PCR method for simultaneous detection and differentiation of COVID-19 and Influenza A/B |
| title_short | Design and assessment of a multiplex real-time PCR method for simultaneous detection and differentiation of COVID-19 and Influenza A/B |
| title_sort | design and assessment of a multiplex real time pcr method for simultaneous detection and differentiation of covid 19 and influenza a b |
| topic | Multiplex; Real-time polymerase chain reaction; SARS-CoV-2; COVID-19; Influenza B; Influenza A |
| url | https://ijm.tums.ac.ir/index.php/ijm/article/view/5047 |
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