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

Seyed Mohammad Amin  Mousavi-Rad; Shohreh Zare  Karizi; Hamid  Sedighian; Seyed Ali  Mirhosseini; Hadi Esmaeili  Gouvarchin ghaleh; Jafar  Amani

doi:10.18502/ijm.v17i3.18832

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

DOI: https://doi.org/10.18502/ijm.v17i3.18832

Keywords: Human papillomavirus; Polymerase chain reaction-Enzyme-linked immunosorbent assay; Molecular diagnos- tics; Hybridization techniques; HPV DNA detection

Abstract

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. Addi- tionally, 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 stan- dard. 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 re- source-limited settings. Its simplicity and compatibility with existing workflows makes it a promising tool for routine diag- nostic applications.