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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Main Authors: 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
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-01-01
Series:PLoS Neglected Tropical Diseases
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.
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publishDate 2025-01-01
publisher Public Library of Science (PLoS)
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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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