Related Experiment Video
Updated: Jun 25, 2026
Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
Published on: March 30, 2015
Point-of-care hrHPV mRNA detection with reverse transcription recombinase polymerase amplification on a portable
Emilie Newsham Novak1, Cannon J Hansen1, Zoha Wazir1
1Department of Bioengineering, Rice University, Houston, TX, USA.
Background:
Cervical cancer is the most common cancer in women in Mozambique, where limited screening infrastructure contributes to high disease burden. While testing for high-risk human papillomavirus (hrHPV) DNA can identify women at risk for cervical cancer, its limited specificity may lead to overtreatment in screen-and-treat programs, straining resources. We performed a pilot study to evaluate preliminary feasibility of a novel, accessible test to detect hrHPV mRNA, a more specific biomarker for cervical cancer risk, from 22 women who previously screened positive for visual inspection with acetic acid at Maputo Central Hospital in Mozambique.
Methods:
Cervicovaginal samples were analyzed for hrHPV DNA using the Xpert HPV test. RNA was extracted from samples using a benchtop spinner instead of a centrifuge. Isothermal reverse transcription recombinase polymerase amplification (RT-RPA) assays targeting HPV 16, 18, and 45 mRNA, plus a cellular control were performed on a portable fluorimeter with results compared to the gold standard portable reverse transcription quantitative polymerase chain reaction (RT-qPCR). Cervical biopsies were obtained and submitted for histopathologic analysis.
Results:
RT-RPA and RT-qPCR results were in 100% agreement (22/22). The sample-to-answer times were 1.5 h for the RT-RPA test and 3.75 h for RT-qPCR. All RT-RPA-positive samples were Xpert-positive for the same HPV type. Two Xpert-positive samples were RT-RPA-negative; four were positive for high-risk HPV types other than 16, 18, or 45. RT-RPA had a higher positive predictive value (100%) for high grade lesions by biopsy than Xpert (73%). Xpert had a higher negative predictive value (82%) than RT-RPA (71%) for high grade lesions.
Conclusions:
This study demonstrates the potential feasibility of novel point-of-care hrHPV mRNA testing in a low-income country. Additional work is needed before this test is clinically relevant and deployable in resource-limited settings.
Trial Registration:
This study was registered on 2022-04-25 on clinicaltrials.gov under registration number NCT05372484.
