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Operational Feasibility and Performance of mRPA Versus PCR for the Detection of HPV Types 16, 18, and 52 in Clinical
Monica Dwi Hartanti1,2, Rahmi Amtha3, Raditya Wratsangka1
1Faculty of Medicine, Universitas Trisakti, West Jakarta, Jakarta, Indonesia.
Purpose:
This exploratory study investigates the feasibility and performance of multiplex Recombinase Polymerase Amplification (mRPA) compared to conventional Polymerase Chain Reaction (PCR) for the detection and genotyping of high-risk Human Papillomavirus (HPV) types 16, 18, and 52. Current PCR methods are widely used for HPV detection but are limited by the need for complex thermal cycling equipment and lengthy processing times, which restrict their use in low-resource settings. This study aims to evaluate whether mRPA can serve as a faster, simpler, and more accessible alternative for HPV screening in primary healthcare environments.
Methods:
A total of 20 clinical samples from cervical swabs were tested using both mRPA and conventional PCR. The samples were preserved in ThinPrep® Specimen Collection fluid and stored at -20°C. mRPA reactions were conducted under isothermal conditions at 39°C for 30 minutes, while conventional PCR followed standard cycling protocols. Sensitivity, specificity, operational efficiency, and feasibility in low-resource settings were assessed and compared between the two methods. The study complies with the Declaration of Helsinki and was approved by the Ethics Committee of Faculty of Medicine, Universitas Indonesia.
Results:
The mRPA demonstrated sensitivity and specificity that were lower than PCR, with detection rates of 100% for HPV 16, 80% for HPV 18, and 60% for HPV 52, compared to PCR's 100% across all types. Overall, mRPA achieved an overall sensitivity of 80% and specificity of 100%. However, mRPA significantly reduced the amplification time to 30 minutes and eliminated the need for thermal cyclers, highlighting its potential suitability for primary healthcare settings. The practical implications of mRPA's rapid turnaround time and simplified equipment requirements make it a promising tool for point-of-care applications in resource-limited environments.
Conclusion:
The findings suggest that mRPA could serve as a viable alternative to conventional PCR for HPV genotyping, offering advantages in speed and simplicity. Although mRPA's diagnostic performance was lower than PCR, its operational benefits make it particularly suitable for use in resource-limited settings. Future research should focus on further optimization and validation to enhance mRPA's diagnostic accuracy and explore its integration with user-friendly detection platforms.
