A Rapid, Sensitive and Type-Specific Detection of High-Risk HPV-16 and HPV-18

Abstract

Human papillomavirus (HPV) infection, particularly infection with HPVs 16 and 18, is a major cause of cervical cancer. The current high-risk HPV screening or diagnosis tests use cytological or molecular techniques that are primarily based on qualitative HPV DNA detection. Comparative studies, however, revealed that different assays have varying sensitivities for detecting specific HPV types. Here, we developed and optimized a sensitive PCR (Polymerase Chain Reaction) assay for detection of high-risk HPV-16 and HPV-18. The PCR parameters were optimized, and analytical specificities were validated. Performance of developed PCR assay was evaluated in clinical samples (n = 100) which showed 100% specificity for both the assays and 96.97% and 94.12% sensitivity for HPV-16 and HPV-18, respectively. The developed assay demonstrated high sensitivity and specificity for detection of high-risk HPV-16 and HPV-18, making it applicable to routine HPV detection practices.

Keywords: Cervical cancer; Detection; Human papillomavirus; PCR.

Fig. 1

PCR optimization for HPV-16. a Annealing Temperature Optimization: Optimum annealing temperature was observed at 56 °C b Primer Concentration Optimization: Optimum primer concentration was observed to be 0.5 µl. c Template Concentration Optimization: Optimum template concentration was observed to be 1 µl

Fig. 2

PCR optimization for HPV-18. a Annealing Temperature Optimization: Optimum annealing temperature was observed at 53.5 °C. b Primer Concentration Optimization: Optimum primer concentration was observed to be 0.5 µl. c Template Concentration Optimization: Optimum template concentration was observed to be 2 µl