Colorimetric reverse transcription loop-mediated isothermal amplification-based approach for early detection of cancer-related pathogens

Sexually transmitted infections are linked to increased risk of some cancers, with chronic inflammation and persistent infections accounting for roughly 20% of cancer cases worldwide. Early detection of these pathogens can help prevent cancer development or progression. This study focused on Human Immunodeficiency Virus-1, Human Papilloma Virus-16, and Epstein-Barr Virus, known to increase cancer risk. A simple colorimetric and reverse transcription Loop-Mediated Isothermal Amplification (LAMP) assay was developed and standardized for rapid detection of these pathogens. The amplification can be visually detected based on color changes. Specifically designed primers targeting conserved genes of HIV-1, EBV, and HPV-16 were analyzed for their specificity and limit of detection. No cross-reactivity was noticed with other pathogenic RNAs and primer sets displayed 100% specificity to their respective pathogenic RNA. We were able to detect as low as 4 RNA copies per reaction within a brief period of 25 minutes. LAMP amplifies a target nucleic acid sequence under isothermal conditions, requiring only basic equipment like a water bath or heating block. Our results show that LAMP’s simplicity and reliability make it ideal for point-of-need (PON) use, with its early detection of cancer-related pathogens offering significant potential for cancer prevention, public health education, and interventions.