مقالات پذیرفته شده در نهمین کنگره بین المللی زیست پزشکی
A Novel Approach in HPV Management: Precise Genotyping via Hybrid PCR and Targeted Therapy Using CRISPR-Cas Systems
A Novel Approach in HPV Management: Precise Genotyping via Hybrid PCR and Targeted Therapy Using CRISPR-Cas Systems
Saghar Modaresi,1,*
1. Independent Researcher, B.Sc. in Laboratory Sciences
Introduction: Human papillomavirus (HPV) is one of the most prevalent sexually transmitted infections worldwide, with over 200 genotypes identified. Among them, high-risk types such as HPV-16 and HPV-18 are the leading causes of cervical and oropharyngeal cancers. HPV is a non-enveloped double-stranded DNA virus with an ~8,000 bp circular genome, comprising early (E1–E7), late (L1–L2), and regulatory (LCR) regions. The E6 and E7 oncoproteins interfere with tumor suppressors (p53 and Rb), leading to uncontrolled proliferation and tumorigenesis. Accurate genotyping is essential for prognosis, treatment selection, and vaccine development. This review highlights advances in HPV genotyping—particularly Hybrid PCR—and targeted therapeutic approaches using CRISPR-Cas systems.
Methods: This review was conducted by systematically searching PubMed, Scopus, and Google Scholar databases for literature published between 2020 and 2024. Keywords included "HPV genotyping," "Hybrid PCR," "CRISPR-Cas," and "HPV treatment." A total of 11 peer-reviewed articles were selected, focusing on diagnostic advances, therapeutic genome editing, and point-of-care technologies. Comparative evaluation of sensitivity, specificity, practicality, and limitations was carried out for each technology
Results: Hybrid PCR, a reverse hybridization technique, enables simultaneous detection of multiple HPV genotypes with high sensitivity (~95–98%). It surpasses conventional PCR in detecting co-infections, though it is limited to pre-designed probes in kits. Real-time PCR is faster but restricted to a few high-risk genotypes. NGS offers unparalleled resolution but is costly.
CRISPR-based assays, particularly CRISPR/Cas12a integrated with isothermal amplification (e.g., RPA), allow rapid, specific, and multiplex detection of HPV in under 1 hour. These point-of-care tests are suitable for low-resource settings. Therapeutically, CRISPR/Cas9 targeting of E6/E7 has shown efficacy in reactivating p53/Rb pathways, inducing apoptosis in HPV-positive cancer cells. Delivery methods and off-target risks remain challenges.
Conclusion: Hybrid PCR offers an effective and precise approach for HPV genotyping, while CRISPR-Cas platforms are transforming both diagnostics and targeted therapies. Their integration may enable personalized and decentralized HPV care. Further clinical validation, safe delivery systems, and scalable implementation are necessary for widespread adoption.