Abstract

Background: Human papillomavirus (HPV) infection is the leading cause of cervical cancer, with infected individuals at significantly higher risk. Early detection of high-risk HPV strains is crucial for effective cancer prevention. Loopmediated isothermal amplification (LAMP) has emerged as a promising alternative to polymerase chain reaction (PCR) for DNA amplification, offering rapid, costeffective, and isothermal amplification without the need for thermal cycling. Objective: This study aimed to develop a point-of-care (POC) method for HPV screening using LAMP, targeting high-risk HPV strains through the amplification of the E6 and E7 genes. The performance of LAMP was compared with PCR and histopathology findings. Methods: LAMP primers were designed targeting conserved regions of high-risk HPV genomes using three bioinformatics tools: Primer Explorer V4, Opti Gene, and Premier Biosoft. Primer selection was based on melting temperature (Tm), GC content, and secondary structure formation. The LAMP assay was optimized for a one-tube reaction, and positive samples were visually detected using a neutral red indicator. Results: Primers designed with Primer Explorer V4 demonstrated the best performance, with optimal Tm values, high specificity, and minimal secondary structure formation. The LAMP assay successfully detected high-risk HPV strains with rapid amplification and visual detection. Compared to PCR and histopathology, LAMP showed high concordance, making it a reliable screening tool. Conclusion: LAMP, when combined with the PrimerExplorer V4 primer design, is an efficient and reliable method for rapid detection of high-risk HPV. This method has the potential to serve as a point-of-care cervical cancer screening tool, particularly in resource-limited settings.

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