Development and validation of a high-throughput qPCR platform for the detection of soil-transmitted helminth infections.
<h4>Background</h4>Historically, soil-transmitted helminth (STH) control and prevention strategies have relied on mass drug administration efforts targeting preschool and school-aged children. While these efforts have succeeded in reducing morbidity associated with STH infection, recent...
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2025-01-01
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| Online Access: | https://doi.org/10.1371/journal.pntd.0012760 |
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| author | Nils Pilotte Victor Omballa Monica Voss Leah Padgett Malathi Manuel Jeanne L Goodman Tim Littlewood Zayina Zondervenni Manoharan Lisette van Lieshout Jaco J Verweij Manigandan Sekar Ajith Kumar Muthukumar Gretchen Walch Abriana Warren Mariyam Sheikh Andrew Gonzalez Sean R Galagan Sitara Swarna Rao Ajjampur Moudachirou Ibikounlé Steven A Williams Doug Rains Ushashi Dadwal Judd L Walson |
| author_facet | Nils Pilotte Victor Omballa Monica Voss Leah Padgett Malathi Manuel Jeanne L Goodman Tim Littlewood Zayina Zondervenni Manoharan Lisette van Lieshout Jaco J Verweij Manigandan Sekar Ajith Kumar Muthukumar Gretchen Walch Abriana Warren Mariyam Sheikh Andrew Gonzalez Sean R Galagan Sitara Swarna Rao Ajjampur Moudachirou Ibikounlé Steven A Williams Doug Rains Ushashi Dadwal Judd L Walson |
| author_sort | Nils Pilotte |
| collection | DOAJ |
| description | <h4>Background</h4>Historically, soil-transmitted helminth (STH) control and prevention strategies have relied on mass drug administration efforts targeting preschool and school-aged children. While these efforts have succeeded in reducing morbidity associated with STH infection, recent modeling efforts have suggested that expanding intervention to treatment of the entire community could achieve transmission interruption in some settings. Testing the feasibility of such an approach requires large-scale clinical trials, such as the DeWorm3 cluster randomized trial. In addition, accurate interpretation of trial outcomes requires diagnostic platforms capable of accurately determining infection prevalence (particularly as infection intensity is reduced) at large population scale and with significant throughput. Here, we describe the development and validation of such a high-throughput molecular testing platform.<h4>Methodology/principal findings</h4>Through the development, selection, and validation of appropriate controls, we have successfully created and evaluated the performance of a testing platform capable of the semi-automated, high-throughput detection of four species of STH in human stool samples. Comparison of this platform with singleplex reference assays for the detection of these same pathogens has demonstrated comparable performance metrics, with index assay accuracy measuring at or above 99.5% and 98.1% for each target species at the level of the technical replicate and individual extraction respectively. Through the implementation of a rigorous validation program, we have developed a diagnostic platform capable of providing the necessary throughput and performance required to meet the needs of the DeWorm3 cluster randomized trial and other large-scale operational research efforts for STH.<h4>Conclusions/significance</h4>Resulting from the rigorous developmental approach taken, the platform we describe here provides the needed confidence in testing outcomes when utilized in conjunction with large-scale efforts such as the DeWorm3 trial. Additionally, the successful development of an evaluation and validation strategy provides a template for the creation of similar diagnostic platforms for other neglected tropical diseases. |
| format | Article |
| id | doaj-art-0ef59ccc611a413dbbb67f26d7e9215d |
| institution | Kabale University |
| issn | 1935-2727 1935-2735 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj-art-0ef59ccc611a413dbbb67f26d7e9215d2025-02-05T05:33:28ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352025-01-01191e001276010.1371/journal.pntd.0012760Development and validation of a high-throughput qPCR platform for the detection of soil-transmitted helminth infections.Nils PilotteVictor OmballaMonica VossLeah PadgettMalathi ManuelJeanne L GoodmanTim LittlewoodZayina Zondervenni ManoharanLisette van LieshoutJaco J VerweijManigandan SekarAjith Kumar MuthukumarGretchen WalchAbriana WarrenMariyam SheikhAndrew GonzalezSean R GalaganSitara Swarna Rao AjjampurMoudachirou IbikounléSteven A WilliamsDoug RainsUshashi DadwalJudd L Walson<h4>Background</h4>Historically, soil-transmitted helminth (STH) control and prevention strategies have relied on mass drug administration efforts targeting preschool and school-aged children. While these efforts have succeeded in reducing morbidity associated with STH infection, recent modeling efforts have suggested that expanding intervention to treatment of the entire community could achieve transmission interruption in some settings. Testing the feasibility of such an approach requires large-scale clinical trials, such as the DeWorm3 cluster randomized trial. In addition, accurate interpretation of trial outcomes requires diagnostic platforms capable of accurately determining infection prevalence (particularly as infection intensity is reduced) at large population scale and with significant throughput. Here, we describe the development and validation of such a high-throughput molecular testing platform.<h4>Methodology/principal findings</h4>Through the development, selection, and validation of appropriate controls, we have successfully created and evaluated the performance of a testing platform capable of the semi-automated, high-throughput detection of four species of STH in human stool samples. Comparison of this platform with singleplex reference assays for the detection of these same pathogens has demonstrated comparable performance metrics, with index assay accuracy measuring at or above 99.5% and 98.1% for each target species at the level of the technical replicate and individual extraction respectively. Through the implementation of a rigorous validation program, we have developed a diagnostic platform capable of providing the necessary throughput and performance required to meet the needs of the DeWorm3 cluster randomized trial and other large-scale operational research efforts for STH.<h4>Conclusions/significance</h4>Resulting from the rigorous developmental approach taken, the platform we describe here provides the needed confidence in testing outcomes when utilized in conjunction with large-scale efforts such as the DeWorm3 trial. Additionally, the successful development of an evaluation and validation strategy provides a template for the creation of similar diagnostic platforms for other neglected tropical diseases.https://doi.org/10.1371/journal.pntd.0012760 |
| spellingShingle | Nils Pilotte Victor Omballa Monica Voss Leah Padgett Malathi Manuel Jeanne L Goodman Tim Littlewood Zayina Zondervenni Manoharan Lisette van Lieshout Jaco J Verweij Manigandan Sekar Ajith Kumar Muthukumar Gretchen Walch Abriana Warren Mariyam Sheikh Andrew Gonzalez Sean R Galagan Sitara Swarna Rao Ajjampur Moudachirou Ibikounlé Steven A Williams Doug Rains Ushashi Dadwal Judd L Walson Development and validation of a high-throughput qPCR platform for the detection of soil-transmitted helminth infections. PLoS Neglected Tropical Diseases |
| title | Development and validation of a high-throughput qPCR platform for the detection of soil-transmitted helminth infections. |
| title_full | Development and validation of a high-throughput qPCR platform for the detection of soil-transmitted helminth infections. |
| title_fullStr | Development and validation of a high-throughput qPCR platform for the detection of soil-transmitted helminth infections. |
| title_full_unstemmed | Development and validation of a high-throughput qPCR platform for the detection of soil-transmitted helminth infections. |
| title_short | Development and validation of a high-throughput qPCR platform for the detection of soil-transmitted helminth infections. |
| title_sort | development and validation of a high throughput qpcr platform for the detection of soil transmitted helminth infections |
| url | https://doi.org/10.1371/journal.pntd.0012760 |
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