Point-of-care hrHPV mRNA detection with reverse transcription recombinase polymerase amplification on a portable fluorimeter from provider-collected cervical samples in Maputo, Mozambique.
Emilie Newsham Novak, Cannon J Hansen, Zoha Wazir, Ariel Ma, Jennifer Carns, Hira Atif, Jane R Montealegre, Michael E Scheurer, Cesaltina Lorenzoni, Daniel G Rosen, Mila P Salcedo, Kathleen M Schmeler, Rebecca R Richards-Kortum
{"title":"Point-of-care hrHPV mRNA detection with reverse transcription recombinase polymerase amplification on a portable fluorimeter from provider-collected cervical samples in Maputo, Mozambique.","authors":"Emilie Newsham Novak, Cannon J Hansen, Zoha Wazir, Ariel Ma, Jennifer Carns, Hira Atif, Jane R Montealegre, Michael E Scheurer, Cesaltina Lorenzoni, Daniel G Rosen, Mila P Salcedo, Kathleen M Schmeler, Rebecca R Richards-Kortum","doi":"10.1186/s13027-026-00772-5","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p><p><strong>Trial registration: </strong>This study was registered on 2022-04-25 on clinicaltrials.gov under registration number NCT05372484.</p>","PeriodicalId":13568,"journal":{"name":"Infectious Agents and Cancer","volume":" ","pages":""},"PeriodicalIF":3.2000,"publicationDate":"2026-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13393838/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Infectious Agents and Cancer","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1186/s13027-026-00772-5","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"IMMUNOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
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.
期刊介绍：
Infectious Agents and Cancer is an open access, peer-reviewed online journal that encompasses all aspects of basic, clinical, epidemiological and translational research providing an insight into the association between chronic infections and cancer.
The journal welcomes submissions in the pathogen-related cancer areas and other related topics, in particular:
• HPV and anogenital cancers, as well as head and neck cancers;
• EBV and Burkitt lymphoma;
• HCV/HBV and hepatocellular carcinoma as well as lymphoproliferative diseases;
• HHV8 and Kaposi sarcoma;
• HTLV and leukemia;
• Cancers in Low- and Middle-income countries.
The link between infection and cancer has become well established over the past 50 years, and infection-associated cancer contribute up to 16% of cancers in developed countries and 33% in less developed countries.
Preventive vaccines have been developed for only two cancer-causing viruses, highlighting both the opportunity to prevent infection-associated cancers by vaccination and the gaps that remain before vaccines can be developed for other cancer-causing agents. These gaps are due to incomplete understanding of the basic biology, natural history, epidemiology of many of the pathogens that cause cancer, the mechanisms they exploit to cause cancer, and how to interrupt progression to cancer in human populations. Early diagnosis or identification of lesions at high risk of progression represent the current most critical research area of the field supported by recent advances in genomics and proteomics technologies.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
