Development of a multiplex quantitative real-time PCR (qPCR) assay for simultaneous detection of Schistosoma haematobium and high-risk human papillomaviruses
9 affiliations
- 1Centre for Research in Applied Biology, University of Energy and Natural Resources, Sunyani, Ghana
- 2Department of Medical Diagnostics, Faculty of Allied Health Science, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana
- 3Department of Chemical Pathology, School of Medical Sciences, University of Cape Coast, Cape Coast, Ghana
- 4Department of Medical Laboratory Science, University of Ghana, Accra, Legon, Ghana
- 5Department of Theoretical and Applied Biology, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana
- 6Dr. K C Patel Research and Development Centre (KRADLE), Charotar University of Science and Technology (CHARUSAT), Changa, India
- 7Department of Population, Family and Reproductive Health, School of Public Health, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana
- 8Department of Biological Science, University of Energy and Natural Resources, Sunyani, Ghana
- 9International HPV Reference Center, Center for Cervical Cancer Elimination, Karolinska Institutet, Stockholm, Sweden
Abstract
Background: Female genital schistosomiasis (FGS) and high-risk human papillomavirus (hr-HPV) infections frequently co-occur in sub-Saharan Africa and may jointly contribute to cervical carcinogenesis. However, no existing molecular platform enables simultaneous detection of Schistosoma haematobium and hr-HPV from a single cervical specimen. Methods: A multiplex quantitative real-time PCR (qPCR) assay targeting the S. haematobium Dra1 repeat and seven hr-HPV genotypes (16, 18, 31, 33, 45, 52, 58) was developed and validated. Analytical performance was assessed using HPV reference plasmids from the International HPV Reference Centre and biobanked S. haematobium DNA from the Centre for Research in Applied Biology (CeRAB), Ghana. Clinical performance was evaluated using 217 archived cervical swabs from high-risk women enrolled in the CERVIVAL project in rural Ghana. Results: The assay showed a limit of detection of 10 copies/µL for all targets, with no cross-reactivity to non-target HPV genotypes, common uropathogenic bacteria, or Schistosoma mansoni. In reference panels, positive and negative agreement were 98.2% and 100% for S. haematobium and 100% for both sensitivity and specificity across all hr-HPV genotypes. In the 217 high-risk cervical samples, S. haematobium DNA was detected in 56.7% (123/217). HPV58 (19.8%), HPV16 (14.7%), and HPV52 (12.4%) were the most frequent genotypes. Concordance with visual inspection/microscopy for FGS was 93.6% (κ = 0.87), and concordance with a molecular HPV test (ScreenFire HPV) ranged from 90.3% to 99.5% (κ = 0.80–0.95) across HPV channels. The assay identified 11 additional S. haematobium–positive samples that were negative by visual inspection/microscopy. Conclusions: The multiplex qPCR assay provides a highly sensitive and practical tool for integrated detection of S. haematobium and hr-HPV from a single cervical sample. Its compatibility with standard qPCR platforms and strong agreement with established diagnostic methods support its potential use in FGS and cervical cancer screening programmes in resource-limited, co-endemic settings. Author summary: In sub-Saharan Africa, millions of women face the dual threat of infections that severely impact reproductive health: a parasitic worm known as Schistosoma haematobium, which causes female genital schistosomiasis, and high-risk types of human papillomavirus, the primary cause of cervical cancer. Growing evidence suggests that suffering from both infections simultaneously may increase a woman’s risk of developing cervical cancer. However, health programs currently lack a simple method to test for both pathogens using a single clinical sample. To address this gap, we developed a new laboratory test capable of detecting DNA from both the parasite and the virus in a single cervical swab. When we evaluated this tool using samples from women in Ghana, we found it to be highly accurate. By combining these diagnoses into one highly sensitive tool that uses standard laboratory equipment, we can streamline screening programs. This approach reduces testing resources and could potentially allow health workers to identify and treat these conditions earlier, offering a practical solution to help prevent cervical cancer in regions where both diseases are widespread.
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BibTeX
@article{donkoh2026development,
title = {{Development of a multiplex quantitative real-time PCR (qPCR) assay for simultaneous detection of Schistosoma haematobium and high-risk human papillomaviruses}},
author = {Emmanuel Timmy Donkoh and Isaac Williams and Rita Nyaaba Akologo and Alfred Afriyie Asiedu and Reindoff Junior Ofori-Atta and Ivy Wina Boadu and Oksana Ryabinina and Akua Obeng Forson and Kwadwo Boampong and Neeraj Jain and Edward Tieru Dassah and Samuel Fosu Gyasi and Laila Sara Arroyo Mühr and William Gariba Akanwariwaik},
journal = {PLOS Neglected Tropical Diseases},
year = {2026},
month = sep,
doi = {10.1371/journal.pntd.0014694},
url = {https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0014694},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Development of a multiplex quantitative real-time PCR (qPCR) assay for simultaneous detection of Schistosoma haematobium and high-risk human papillomaviruses
AU - Emmanuel Timmy Donkoh
AU - Isaac Williams
AU - Rita Nyaaba Akologo
AU - Alfred Afriyie Asiedu
AU - Reindoff Junior Ofori-Atta
AU - Ivy Wina Boadu
AU - Oksana Ryabinina
AU - Akua Obeng Forson
AU - Kwadwo Boampong
AU - Neeraj Jain
AU - Edward Tieru Dassah
AU - Samuel Fosu Gyasi
AU - Laila Sara Arroyo Mühr
AU - William Gariba Akanwariwaik
PY - 2026
DA - 2026/09/11
JO - PLOS Neglected Tropical Diseases
DO - 10.1371/journal.pntd.0014694
UR - https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0014694
AB - Background: Female genital schistosomiasis (FGS) and high-risk human papillomavirus (hr-HPV) infections frequently co-occur in sub-Saharan Africa and may jointly contribute to cervical carcinogenesis. However, no existing molecular platform enables simultaneous detection of Schistosoma haematobium and hr-HPV from a single cervical specimen. Methods: A multiplex quantitative real-time PCR (qPCR) assay targeting the S. haematobium Dra1 repeat and seven hr-HPV genotypes (16, 18, 31, 33, 45, 52, 58) was developed and validated. Analytical performance was assessed using HPV reference plasmids from the International HPV Reference Centre and biobanked S. haematobium DNA from the Centre for Research in Applied Biology (CeRAB), Ghana. Clinical performance was evaluated using 217 archived cervical swabs from high-risk women enrolled in the CERVIVAL project in rural Ghana. Results: The assay showed a limit of detection of 10 copies/µL for all targets, with no cross-reactivity to non-target HPV genotypes, common uropathogenic bacteria, or Schistosoma mansoni. In reference panels, positive and negative agreement were 98.2% and 100% for S. haematobium and 100% for both sensitivity and specificity across all hr-HPV genotypes. In the 217 high-risk cervical samples, S. haematobium DNA was detected in 56.7% (123/217). HPV58 (19.8%), HPV16 (14.7%), and HPV52 (12.4%) were the most frequent genotypes. Concordance with visual inspection/microscopy for FGS was 93.6% (κ = 0.87), and concordance with a molecular HPV test (ScreenFire HPV) ranged from 90.3% to 99.5% (κ = 0.80–0.95) across HPV channels. The assay identified 11 additional S. haematobium–positive samples that were negative by visual inspection/microscopy. Conclusions: The multiplex qPCR assay provides a highly sensitive and practical tool for integrated detection of S. haematobium and hr-HPV from a single cervical sample. Its compatibility with standard qPCR platforms and strong agreement with established diagnostic methods support its potential use in FGS and cervical cancer screening programmes in resource-limited, co-endemic settings. Author summary: In sub-Saharan Africa, millions of women face the dual threat of infections that severely impact reproductive health: a parasitic worm known as Schistosoma haematobium, which causes female genital schistosomiasis, and high-risk types of human papillomavirus, the primary cause of cervical cancer. Growing evidence suggests that suffering from both infections simultaneously may increase a woman’s risk of developing cervical cancer. However, health programs currently lack a simple method to test for both pathogens using a single clinical sample. To address this gap, we developed a new laboratory test capable of detecting DNA from both the parasite and the virus in a single cervical swab. When we evaluated this tool using samples from women in Ghana, we found it to be highly accurate. By combining these diagnoses into one highly sensitive tool that uses standard laboratory equipment, we can streamline screening programs. This approach reduces testing resources and could potentially allow health workers to identify and treat these conditions earlier, offering a practical solution to help prevent cervical cancer in regions where both diseases are widespread.
SN - 1935-2735
ER -