Hartanti Monica Dwi, Amtha Rahmi, Wratsangka Raditya, Rinanti Astri, Djuana Endang, Hizbulloh Ilham, Teguh Christopher Andrew, Kogoya Alvionita, Subekti Didik T, Desem Muhammad Ibrahim, Lestari Christina Safira Whinie, Asmaria Talitha
Faculty of Medicine, Universitas Trisakti, West Jakarta, Jakarta, Indonesia.
Center for Biomedical Research, Research Organization for Health, National Research and Innovation Agency (BRIN), Cibinong, West Java, Indonesia.
Int J Womens Health. 2025 Apr 5;17:955-964. doi: 10.2147/IJWH.S496621. eCollection 2025.
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.
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.
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.
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.
本探索性研究调查多重重组酶聚合酶扩增（mRPA）与传统聚合酶链反应（PCR）相比，用于检测高危人乳头瘤病毒（HPV）16、18和52型并进行基因分型的可行性和性能。当前的PCR方法广泛用于HPV检测，但受限于需要复杂的热循环设备和较长的处理时间，这限制了它们在资源匮乏环境中的使用。本研究旨在评估mRPA是否可作为基层医疗环境中HPV筛查的一种更快、更简单且更易获得的替代方法。
使用mRPA和传统PCR对总共20份宫颈拭子临床样本进行检测。样本保存在ThinPrep样本采集液中，并储存在-20°C。mRPA反应在39°C等温条件下进行30分钟，而传统PCR遵循标准循环方案。评估并比较了两种方法在低资源环境中的敏感性、特异性、操作效率和可行性。本研究遵循《赫尔辛基宣言》，并获得印度尼西亚大学医学院伦理委员会的批准。
mRPA的敏感性和特异性低于PCR，HPV 16型的检测率为100%，HPV 18型为80%，HPV 52型为60%，而PCR对所有类型的检测率均为100%。总体而言，mRPA的总体敏感性为80%，特异性为100%。然而，mRPA显著缩短了扩增时间至30分钟，且无需热循环仪，凸显了其在基层医疗环境中的潜在适用性。mRPA快速周转时间和简化设备要求的实际意义使其成为资源有限环境中即时检测应用的有前景工具。
研究结果表明，mRPA可作为传统PCR进行HPV基因分型的可行替代方法，在速度和简便性方面具有优势。尽管mRPA的诊断性能低于PCR，但其操作优势使其特别适合在资源有限的环境中使用。未来的研究应专注于进一步优化和验证，以提高mRPA的诊断准确性，并探索其与用户友好型检测平台的整合。
