Successful Retrieval of Human Papillomavirus DNA in Veil-Based Collected Female Genital Secretions After Long-Term Storage in Universal Transport Medium
<b>Background/Objectives</b>: 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 molecul...
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MDPI AG
2025-04-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/15/9/1079 |
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| author | Jonathan Muwonga Tukisadila Juval Avala Ntsigouaye Serge Tonen-Wolyec Ralph-Sydney Mboumba Bouassa Jeremie Muwonga Laurent Belec |
| author_facet | Jonathan Muwonga Tukisadila Juval Avala Ntsigouaye Serge Tonen-Wolyec Ralph-Sydney Mboumba Bouassa Jeremie Muwonga Laurent Belec |
| author_sort | Jonathan Muwonga Tukisadila |
| collection | DOAJ |
| description | <b>Background/Objectives</b>: 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. <b>Methods</b>: The detection and quantification of HPV DNA were evaluated in female genital secretions self-collected using veils and conserved in Cyt-All<sup>®</sup> UTM at −30 °C, +4 °C, and +25 °C after long-term 27-month storage. <b>Results</b>: 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<sup>®</sup> medium, even at room temperature, allows for the detection and quantification of any HPV and HR-HPV with high accuracy. <b>Conclusions</b>: 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. |
| format | Article |
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| institution | OA Journals |
| issn | 2075-4418 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | MDPI AG |
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| series | Diagnostics |
| spelling | doaj-art-8df8b10fe82e476dafdcbe7b3b61d37b2025-08-20T01:49:14ZengMDPI AGDiagnostics2075-44182025-04-01159107910.3390/diagnostics15091079Successful Retrieval of Human Papillomavirus DNA in Veil-Based Collected Female Genital Secretions After Long-Term Storage in Universal Transport MediumJonathan Muwonga Tukisadila0Juval Avala Ntsigouaye1Serge Tonen-Wolyec2Ralph-Sydney Mboumba Bouassa3Jeremie Muwonga4Laurent Belec5École Doctorale Régionale D’Afrique Centrale en Infectiologie Tropicale, Franceville 876, GabonÉcole Doctorale Régionale D’Afrique Centrale en Infectiologie Tropicale, Franceville 876, GabonÉcole Doctorale Régionale D’Afrique Centrale en Infectiologie Tropicale, Franceville 876, GabonÉcole Doctorale Régionale D’Afrique Centrale en Infectiologie Tropicale, Franceville 876, GabonLaboratoire de Biologie Clinique des Cliniques Universitaires de Kinshasa, Kinshasa 123, Democratic Republic of the CongoLaboratory of Virology, Hôpital Européen Georges Pompidou, Assistance Publique-Hôpitaux de Paris (AP-HP), 75015 Paris, France<b>Background/Objectives</b>: 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. <b>Methods</b>: The detection and quantification of HPV DNA were evaluated in female genital secretions self-collected using veils and conserved in Cyt-All<sup>®</sup> UTM at −30 °C, +4 °C, and +25 °C after long-term 27-month storage. <b>Results</b>: 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<sup>®</sup> medium, even at room temperature, allows for the detection and quantification of any HPV and HR-HPV with high accuracy. <b>Conclusions</b>: 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.https://www.mdpi.com/2075-4418/15/9/1079human papillomavirus (HPV) DNA preservationself-sampling-based HPV screeninggenital veilmultiplex real-time PCRHPV viral loadbiobanking |
| spellingShingle | Jonathan Muwonga Tukisadila Juval Avala Ntsigouaye Serge Tonen-Wolyec Ralph-Sydney Mboumba Bouassa Jeremie Muwonga Laurent Belec Successful Retrieval of Human Papillomavirus DNA in Veil-Based Collected Female Genital Secretions After Long-Term Storage in Universal Transport Medium Diagnostics human papillomavirus (HPV) DNA preservation self-sampling-based HPV screening genital veil multiplex real-time PCR HPV viral load biobanking |
| title | Successful Retrieval of Human Papillomavirus DNA in Veil-Based Collected Female Genital Secretions After Long-Term Storage in Universal Transport Medium |
| title_full | Successful Retrieval of Human Papillomavirus DNA in Veil-Based Collected Female Genital Secretions After Long-Term Storage in Universal Transport Medium |
| title_fullStr | Successful Retrieval of Human Papillomavirus DNA in Veil-Based Collected Female Genital Secretions After Long-Term Storage in Universal Transport Medium |
| title_full_unstemmed | Successful Retrieval of Human Papillomavirus DNA in Veil-Based Collected Female Genital Secretions After Long-Term Storage in Universal Transport Medium |
| title_short | Successful Retrieval of Human Papillomavirus DNA in Veil-Based Collected Female Genital Secretions After Long-Term Storage in Universal Transport Medium |
| title_sort | successful retrieval of human papillomavirus dna in veil based collected female genital secretions after long term storage in universal transport medium |
| topic | human papillomavirus (HPV) DNA preservation self-sampling-based HPV screening genital veil multiplex real-time PCR HPV viral load biobanking |
| url | https://www.mdpi.com/2075-4418/15/9/1079 |
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