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Design and comparison of PCR-ELISA reaction with other available hybridization methods to identify types 11, 16, and
Seyed Mohammad Amin Mousavi-Rad1, Shohreh Zare Karizi2, Hamid Sedighian1
1Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.
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.
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