Successful Retrieval of Human Papillomavirus DNA in Veil-Based Collected Female Genital Secretions After Long-Term Storage in Universal Transport Medium

Jonathan Muwonga Tukisadila; Juval Avala Ntsigouaye; Serge Tonen-Wolyec; Ralph-Sydney Mboumba Bouassa; Jeremie Muwonga; Laurent Belec

doi:10.3390/diagnostics15091079

Successful Retrieval of Human Papillomavirus DNA in Veil-Based Collected Female Genital Secretions After Long-Term Storage in Universal Transport Medium

Diagnostics (Basel). 2025 Apr 24;15(9):1079. doi: 10.3390/diagnostics15091079.

Authors

Jonathan Muwonga Tukisadila^{1

2

3}, Juval Avala Ntsigouaye¹, Serge Tonen-Wolyec^{1

4

5}, Ralph-Sydney Mboumba Bouassa^{1

6

7}, Jeremie Muwonga², Laurent Belec^{3

8}

Affiliations

¹ École Doctorale Régionale D'Afrique Centrale en Infectiologie Tropicale, Franceville 876, Gabon.
² Laboratoire de Biologie Clinique des Cliniques Universitaires de Kinshasa, Kinshasa 123, Democratic Republic of the Congo.
³ Laboratory of Virology, Hôpital Européen Georges Pompidou, Assistance Publique-Hôpitaux de Paris (AP-HP), 75015 Paris, France.
⁴ Department of Internal Medicine, Faculté de Médecine, Université de Bunia, Bunia 292, Democratic Republic of the Congo.
⁵ Department of Internal Medicine, Faculté de Médecine et Pharmacie, Université de Kisangani, Kisangani 2012, Democratic Republic of the Congo.
⁶ Institut du Savoir Montfort, Montfort Hospital, Ottawa, ON K1K 0T2, Canada.
⁷ Department of Family Medicine, Faculty of Medicine, University of Ottawa, Ottawa, ON K1N 6S1, Canada.
⁸ Faculté de Médecine Paris Descartes, Université Paris Cité, 75006 Paris, France.

Abstract

Background/Objectives: The surveillance of viral strain evolution is needed during prophylactic HPV vaccination programs against cervical cancer and necessitates safely archiving and storing cervical samples while maintaining the long-term stability of HPV DNA to carry out molecular diagnosis. The present proof-of-concept study aimed to assess DNA stability for HPV molecular detection from veils resuspended in a universal transport medium (UTM) and conserved at different temperatures after long-term storage. Methods: The detection and quantification of HPV DNA were evaluated in female genital secretions self-collected using veils and conserved in Cyt-All^® UTM at -30 °C, +4 °C, and +25 °C after long-term 27-month storage. Results: A slight degradation of the ubiquitous single-copy cellular DNA TOP3 gene was assessed using multiplex real-time PCR (BMRT Human Papillomavirus Genotyping Real Time PCR Kit, Bioperfectus Technologies Co., Ltd., Taizhou, Jiangsu, China) at positive temperatures (+4 °C and +25 °C) but not at a frozen temperature (-30 °C) after 27 months of storage. Nevertheless, HPV DNA preservation was sufficient at the three storage temperatures to detect and quantify HPV DNA, with a similar rate of HPV detection, a similar level of cumulative HPV viral loads, high sensitivity and specificity, and perfect concordance in HPV genotype detection after the long period of 27 months of storage. Finally, the conservation of genital samples for a prolonged period in the Cyt-All^® medium, even at room temperature, allows for the detection and quantification of any HPV and HR-HPV with high accuracy. Conclusions: The combination of veil-based self-sampling of female genital secretions and their elution and conservation in UTM may be used in the field to carry out longitudinal molecular epidemiology surveys of circulating HPV.

Keywords: HPV viral load; biobanking; genital veil; human papillomavirus (HPV) DNA preservation; multiplex real-time PCR; self-sampling-based HPV screening; sub-Saharan Africa; universal transport medium.