Development of a simple, rapid, and sensitive molecular diagnostic assay for cholera.
Cholera continues to inflict high rates of morbidity and mortality. Prompt identification of cholera cases facilitates rapid outbreak responses in the short term while providing reliable surveillance data to guide long-term policies and interventions. Microbiological stool culture, the current recog...
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ftdoajarticles:oai:doaj.org/article:f8f03dd69b004f5d88d30b4a7f05c720 2023-05-15T15:12:30+02:00 Development of a simple, rapid, and sensitive molecular diagnostic assay for cholera. Subhra Chakraborty Mirza Velagic Sean Connor 2023-02-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0011113 https://doaj.org/article/f8f03dd69b004f5d88d30b4a7f05c720 EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0011113 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0011113 https://doaj.org/article/f8f03dd69b004f5d88d30b4a7f05c720 PLoS Neglected Tropical Diseases, Vol 17, Iss 2, p e0011113 (2023) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2023 ftdoajarticles https://doi.org/10.1371/journal.pntd.0011113 2023-03-05T01:33:15Z Cholera continues to inflict high rates of morbidity and mortality. Prompt identification of cholera cases facilitates rapid outbreak responses in the short term while providing reliable surveillance data to guide long-term policies and interventions. Microbiological stool culture, the current recognized gold standard for diagnosing cholera, has significant limitations. Rapid diagnostic tests (RDTs) represent promising alternatives for diagnosing cholera in areas with limited laboratory infrastructure. However, studies conducted with the current cholera RDTs demonstrated wide variations in sensitivity and specificity. To address this gap in the diagnosis of cholera, we developed a simple, rapid, and sensitive diagnostic assay, "Rapid LAMP based Diagnostic Test (RLDT)." With a novel, simple sample preparation method directly from the fecal samples along with lyophilized reaction strips and using established Loop-mediated Isothermal Amplification (LAMP) platform, cholera toxin gene (ctxA) and O1 (O1rfb) gene could be detected in less than an hour. Cholera RLDT assay is cold chain and electricity-free. To avoid any end-user bias, a battery-operated, handheld reader was used to read the RLDT results. The performance specifications of the cholera RLDT assay, including analytical sensitivity and specificity, were evaluated using direct fecal samples, dried fecal samples on filter paper, and environmental water samples spiked with cholera strain. The limit of detection (LOD) was ~104 CFU/gm of stool for both ctxA and O1 genes, corresponding to about 1 CFU of Vibrio cholerae per reaction within 40 minutes. The LOD was 10 bacteria per ml of environmental water when tested with RLDT directly, without enrichment. Being simple, RLDT has the potential to be applied in resource-poor endemic settings for rapid, sensitive, and reliable diagnosis of cholera. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 17 2 e0011113 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Subhra Chakraborty Mirza Velagic Sean Connor Development of a simple, rapid, and sensitive molecular diagnostic assay for cholera. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Cholera continues to inflict high rates of morbidity and mortality. Prompt identification of cholera cases facilitates rapid outbreak responses in the short term while providing reliable surveillance data to guide long-term policies and interventions. Microbiological stool culture, the current recognized gold standard for diagnosing cholera, has significant limitations. Rapid diagnostic tests (RDTs) represent promising alternatives for diagnosing cholera in areas with limited laboratory infrastructure. However, studies conducted with the current cholera RDTs demonstrated wide variations in sensitivity and specificity. To address this gap in the diagnosis of cholera, we developed a simple, rapid, and sensitive diagnostic assay, "Rapid LAMP based Diagnostic Test (RLDT)." With a novel, simple sample preparation method directly from the fecal samples along with lyophilized reaction strips and using established Loop-mediated Isothermal Amplification (LAMP) platform, cholera toxin gene (ctxA) and O1 (O1rfb) gene could be detected in less than an hour. Cholera RLDT assay is cold chain and electricity-free. To avoid any end-user bias, a battery-operated, handheld reader was used to read the RLDT results. The performance specifications of the cholera RLDT assay, including analytical sensitivity and specificity, were evaluated using direct fecal samples, dried fecal samples on filter paper, and environmental water samples spiked with cholera strain. The limit of detection (LOD) was ~104 CFU/gm of stool for both ctxA and O1 genes, corresponding to about 1 CFU of Vibrio cholerae per reaction within 40 minutes. The LOD was 10 bacteria per ml of environmental water when tested with RLDT directly, without enrichment. Being simple, RLDT has the potential to be applied in resource-poor endemic settings for rapid, sensitive, and reliable diagnosis of cholera. |
format |
Article in Journal/Newspaper |
author |
Subhra Chakraborty Mirza Velagic Sean Connor |
author_facet |
Subhra Chakraborty Mirza Velagic Sean Connor |
author_sort |
Subhra Chakraborty |
title |
Development of a simple, rapid, and sensitive molecular diagnostic assay for cholera. |
title_short |
Development of a simple, rapid, and sensitive molecular diagnostic assay for cholera. |
title_full |
Development of a simple, rapid, and sensitive molecular diagnostic assay for cholera. |
title_fullStr |
Development of a simple, rapid, and sensitive molecular diagnostic assay for cholera. |
title_full_unstemmed |
Development of a simple, rapid, and sensitive molecular diagnostic assay for cholera. |
title_sort |
development of a simple, rapid, and sensitive molecular diagnostic assay for cholera. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2023 |
url |
https://doi.org/10.1371/journal.pntd.0011113 https://doaj.org/article/f8f03dd69b004f5d88d30b4a7f05c720 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 17, Iss 2, p e0011113 (2023) |
op_relation |
https://doi.org/10.1371/journal.pntd.0011113 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0011113 https://doaj.org/article/f8f03dd69b004f5d88d30b4a7f05c720 |
op_doi |
https://doi.org/10.1371/journal.pntd.0011113 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
17 |
container_issue |
2 |
container_start_page |
e0011113 |
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1766343167668387840 |