Design and comparison of PCR-ELISA reaction with other available hybridization methods to identify types 11, 16, and 18 of the human papillomavirus

Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 17 3 2025 06 02 Design and comparison of PCR-ELISA reaction with other available hybridization methods to identify types 11, 16, and 18 of the human papillomavirus 488 502 Seyed Mohammad Amin Mousavi-Rad Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran Shohreh Zare Karizi Department of Biology, Faculty Member, Varamin-Pishva Branch of Azad University, Tehran, Iran Hamid Sedighian Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran Seyed Ali Mirhosseini Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran Hadi Esmaeili Gouvarchin ghaleh Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Jafar Amani Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran 2024 11 27 2025 01 12 Background and Objectives: Human papillomavirus (HPV) is a significant etiological agent in cervical cancer. This study aimed to evaluate the performance of PCR-ELISA for detecting HPV genotypes 11, 16, and 18 compared to the conventional hybridization methods. Materials and Methods: PCR-ELISA was designed and optimized to detect target HPV genotypes using biotin-labeled probes. Sensitivity, specificity and reproducibility were assessed through intra-assay and inter-assay variability tests. Additionally, a cost-benefit analysis was performed to compare PCR-ELISA with RT-PCR and gel electrophoresis. Results: PCR-ELISA demonstrated high sensitivity (HPV18: 94.92%, HPV16: 98.36%, HPV11: 93.75%) and specificity (100% for all genotypes), with Kappa values ranging from 0.84 to 0.92, indicating strong agreement with the reference standard. Reproducibility analysis showed intra-assay CVs below 5% for most samples and inter-assay CVs within acceptable limits. The cost-benefit analysis revealed significant reductions in reagent and equipment costs compared to RT-PCR, making PCR-ELISA a cost-effective alternative. Conclusion: PCR-ELISA offers a reliable, sensitive and cost-effective method for HPV detection, particularly in resource-limited settings. Its simplicity and compatibility with existing workflows makes it a promising tool for routine diagnostic applications. https://ijm.tums.ac.ir/index.php/ijm/article/view/5156 https://ijm.tums.ac.ir/index.php/ijm/article/download/5156/1791