Fotros Nafiseh, Bashiri Reihaneh, Mohammadi-Yeganeh Samira, Paryan Mahdi
Department of Biotechnology, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
Medical Nanotechnology and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Iran J Microbiol. 2025 Apr;17(2):204-210. doi: 10.18502/ijm.v17i2.18381.
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.
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.
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%.
This multiplex real-time PCR method can accurately detect and distinguish SARS-CoV-2, Influenza B, and Influenza A infections.
呼吸系统病毒感染因其易于传播、具有大流行潜力和显著死亡率，成为一个重大公共问题。诊断这些感染需要实验室检测，因为仅靠临床症状往往并不足够。甲型流感、乙型流感和新冠病毒感染是困扰人群的常见感染，尤其是在冬季。我们开发了一种多重实时聚合酶链反应（PCR）方法，可同时检测甲型流感、乙型流感以及新冠病毒。与现有的检测试剂盒相比，它具有更高的准确性、更低的成本和更快的检测结果，使其成为一种高效的诊断工具。
我们针对甲型流感的M2基因、严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的N基因以及乙型流感的NS1基因设计了引物/TaqMan探针。优化了反应成分，并使用具有已知病毒拷贝数的标准样本测试了功能参数。
该方法对甲型流感和乙型流感的检测限为10拷贝，对SARS-CoV-2为60拷贝。甲型流感的灵敏度和特异性分别为88%和100%，乙型流感为95.6%和100%，SARS-CoV-2为90.4%和100%。
这种多重实时PCR方法能够准确检测并区分SARS-CoV-2、乙型流感和甲型流感感染。
