A novel high throughput assay for anthelmintic drug screening and resistance diagnosis by real-time monitoring of parasite motility.
BACKGROUND: Helminth parasites cause untold morbidity and mortality to billions of people and livestock. Anthelmintic drugs are available but resistance is a problem in livestock parasites, and is a looming threat for human helminths. Testing the efficacy of available anthelmintic drugs and developm...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2010-01-01
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| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | http://europepmc.org/articles/PMC2982823?pdf=render |
| _version_ | 1819144473081282560 |
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| author | Michael J Smout Andrew C Kotze James S McCarthy Alex Loukas |
| author_facet | Michael J Smout Andrew C Kotze James S McCarthy Alex Loukas |
| author_sort | Michael J Smout |
| collection | DOAJ |
| description | BACKGROUND: Helminth parasites cause untold morbidity and mortality to billions of people and livestock. Anthelmintic drugs are available but resistance is a problem in livestock parasites, and is a looming threat for human helminths. Testing the efficacy of available anthelmintic drugs and development of new drugs is hindered by the lack of objective high-throughput screening methods. Currently, drug effect is assessed by observing motility or development of parasites using laborious, subjective, low-throughput methods. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe a novel application for a real-time cell monitoring device (xCELLigence) that can simply and objectively assess anthelmintic effects by measuring parasite motility in real time in a fully automated high-throughput fashion. We quantitatively assessed motility and determined real time IC(50) values of different anthelmintic drugs against several developmental stages of major helminth pathogens of humans and livestock, including larval Haemonchus contortus and Strongyloides ratti, and adult hookworms and blood flukes. The assay enabled quantification of the onset of egg hatching in real time, and the impact of drugs on hatch rate, as well as discriminating between the effects of drugs on motility of drug-susceptible and -resistant isolates of H. contortus. CONCLUSIONS/SIGNIFICANCE: Our findings indicate that this technique will be suitable for discovery and development of new anthelmintic drugs as well as for detection of phenotypic resistance to existing drugs for the majority of helminths and other pathogens where motility is a measure of pathogen viability. The method is also amenable to use for other purposes where motility is assessed, such as gene silencing or antibody-mediated killing. |
| first_indexed | 2024-12-22T12:42:41Z |
| format | Article |
| id | doaj.art-519c2f48f2094aa8ade4679af6ae05cb |
| institution | Directory Open Access Journal |
| issn | 1935-2735 |
| language | English |
| last_indexed | 2024-12-22T12:42:41Z |
| publishDate | 2010-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-519c2f48f2094aa8ade4679af6ae05cb2022-12-21T18:25:24ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27352010-01-01411e88510.1371/journal.pntd.0000885A novel high throughput assay for anthelmintic drug screening and resistance diagnosis by real-time monitoring of parasite motility.Michael J SmoutAndrew C KotzeJames S McCarthyAlex LoukasBACKGROUND: Helminth parasites cause untold morbidity and mortality to billions of people and livestock. Anthelmintic drugs are available but resistance is a problem in livestock parasites, and is a looming threat for human helminths. Testing the efficacy of available anthelmintic drugs and development of new drugs is hindered by the lack of objective high-throughput screening methods. Currently, drug effect is assessed by observing motility or development of parasites using laborious, subjective, low-throughput methods. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe a novel application for a real-time cell monitoring device (xCELLigence) that can simply and objectively assess anthelmintic effects by measuring parasite motility in real time in a fully automated high-throughput fashion. We quantitatively assessed motility and determined real time IC(50) values of different anthelmintic drugs against several developmental stages of major helminth pathogens of humans and livestock, including larval Haemonchus contortus and Strongyloides ratti, and adult hookworms and blood flukes. The assay enabled quantification of the onset of egg hatching in real time, and the impact of drugs on hatch rate, as well as discriminating between the effects of drugs on motility of drug-susceptible and -resistant isolates of H. contortus. CONCLUSIONS/SIGNIFICANCE: Our findings indicate that this technique will be suitable for discovery and development of new anthelmintic drugs as well as for detection of phenotypic resistance to existing drugs for the majority of helminths and other pathogens where motility is a measure of pathogen viability. The method is also amenable to use for other purposes where motility is assessed, such as gene silencing or antibody-mediated killing.http://europepmc.org/articles/PMC2982823?pdf=render |
| spellingShingle | Michael J Smout Andrew C Kotze James S McCarthy Alex Loukas A novel high throughput assay for anthelmintic drug screening and resistance diagnosis by real-time monitoring of parasite motility. PLoS Neglected Tropical Diseases |
| title | A novel high throughput assay for anthelmintic drug screening and resistance diagnosis by real-time monitoring of parasite motility. |
| title_full | A novel high throughput assay for anthelmintic drug screening and resistance diagnosis by real-time monitoring of parasite motility. |
| title_fullStr | A novel high throughput assay for anthelmintic drug screening and resistance diagnosis by real-time monitoring of parasite motility. |
| title_full_unstemmed | A novel high throughput assay for anthelmintic drug screening and resistance diagnosis by real-time monitoring of parasite motility. |
| title_short | A novel high throughput assay for anthelmintic drug screening and resistance diagnosis by real-time monitoring of parasite motility. |
| title_sort | novel high throughput assay for anthelmintic drug screening and resistance diagnosis by real time monitoring of parasite motility |
| url | http://europepmc.org/articles/PMC2982823?pdf=render |
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