Evaluation in Cameroon of a Novel, Simplified Methodology to Assist Molecular Microbiological Analysis of V. cholerae in Resource-Limited Settings.
BACKGROUND:Vibrio cholerae is endemic in South Asia and Africa where outbreaks of cholera occur widely and are particularly associated with poverty and poor sanitation. Knowledge of the genetic diversity of toxigenic V. cholerae isolates, particularly in Africa, remains scarce. The constraints in im...
Published in: | PLOS Neglected Tropical Diseases |
---|---|
Main Authors: | , , , , , , , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
Public Library of Science (PLoS)
2016
|
Subjects: | |
Online Access: | https://doi.org/10.1371/journal.pntd.0004307 https://doaj.org/article/ab92b3cd8dd24d39ae851ace1601b91c |
id |
ftdoajarticles:oai:doaj.org/article:ab92b3cd8dd24d39ae851ace1601b91c |
---|---|
record_format |
openpolar |
spelling |
ftdoajarticles:oai:doaj.org/article:ab92b3cd8dd24d39ae851ace1601b91c 2023-05-15T15:16:48+02:00 Evaluation in Cameroon of a Novel, Simplified Methodology to Assist Molecular Microbiological Analysis of V. cholerae in Resource-Limited Settings. Amanda K Debes Jerome Ateudjieu Etienne Guenou Anna Lena Lopez Mark Philip Bugayong Pearl Joy Retiban Marcelino Garrine Inacio Mandomando Shan Li O Colin Stine David A Sack 2016-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0004307 https://doaj.org/article/ab92b3cd8dd24d39ae851ace1601b91c EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC4703203?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0004307 https://doaj.org/article/ab92b3cd8dd24d39ae851ace1601b91c PLoS Neglected Tropical Diseases, Vol 10, Iss 1, p e0004307 (2016) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2016 ftdoajarticles https://doi.org/10.1371/journal.pntd.0004307 2022-12-30T23:25:26Z BACKGROUND:Vibrio cholerae is endemic in South Asia and Africa where outbreaks of cholera occur widely and are particularly associated with poverty and poor sanitation. Knowledge of the genetic diversity of toxigenic V. cholerae isolates, particularly in Africa, remains scarce. The constraints in improving this understanding is not only the lack of regular cholera disease surveillance, but also the lack of laboratory capabilities in endemic countries to preserve, store and ship isolates in a timely manner. We evaluated the use of simplified sample preservation methods for molecular characterization using multi-locus variable-number tandem-repeat analysis (MLVA) for differentiation of Vibrio cholerae genotypes. METHODS AND FINDINGS:Forty-seven V. cholerae isolates and 18 enriched clinical specimens (e.g. stool specimens after enrichment in broth) from cholera outbreaks in Cameroon were preserved on Whatman filter paper for DNA extraction. The samples were collected from two geographically distinct outbreaks in the Far North of Cameroon (FNC) in June 2014 and October 2014. In addition, a convenience sample of 14 isolates from the Philippines and 8 from Mozambique were analyzed. All 87 DNAs were successfully analyzed including 16 paired samples, one a cultured isolate and the other the enriched specimen from which the isolate was collected. Genotypic results were identical between 15 enriched specimens and their culture isolates and the other pair differed at single locus. Two closely related, but distinct clonal complexes were identified among the Cameroonian specimens from 2014. CONCLUSIONS:Collecting V. cholerae using simplified laboratory methods in remote and low-resource settings allows for subsequent advanced molecular characterization of V. cholerae O1. These simplified DNA preservation methods identify V. cholerae and make possible timely information regarding the genetic diversity of V. cholerae; our results set the stage for continued molecular epidemiological research to better understand the ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 10 1 e0004307 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Amanda K Debes Jerome Ateudjieu Etienne Guenou Anna Lena Lopez Mark Philip Bugayong Pearl Joy Retiban Marcelino Garrine Inacio Mandomando Shan Li O Colin Stine David A Sack Evaluation in Cameroon of a Novel, Simplified Methodology to Assist Molecular Microbiological Analysis of V. cholerae in Resource-Limited Settings. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
BACKGROUND:Vibrio cholerae is endemic in South Asia and Africa where outbreaks of cholera occur widely and are particularly associated with poverty and poor sanitation. Knowledge of the genetic diversity of toxigenic V. cholerae isolates, particularly in Africa, remains scarce. The constraints in improving this understanding is not only the lack of regular cholera disease surveillance, but also the lack of laboratory capabilities in endemic countries to preserve, store and ship isolates in a timely manner. We evaluated the use of simplified sample preservation methods for molecular characterization using multi-locus variable-number tandem-repeat analysis (MLVA) for differentiation of Vibrio cholerae genotypes. METHODS AND FINDINGS:Forty-seven V. cholerae isolates and 18 enriched clinical specimens (e.g. stool specimens after enrichment in broth) from cholera outbreaks in Cameroon were preserved on Whatman filter paper for DNA extraction. The samples were collected from two geographically distinct outbreaks in the Far North of Cameroon (FNC) in June 2014 and October 2014. In addition, a convenience sample of 14 isolates from the Philippines and 8 from Mozambique were analyzed. All 87 DNAs were successfully analyzed including 16 paired samples, one a cultured isolate and the other the enriched specimen from which the isolate was collected. Genotypic results were identical between 15 enriched specimens and their culture isolates and the other pair differed at single locus. Two closely related, but distinct clonal complexes were identified among the Cameroonian specimens from 2014. CONCLUSIONS:Collecting V. cholerae using simplified laboratory methods in remote and low-resource settings allows for subsequent advanced molecular characterization of V. cholerae O1. These simplified DNA preservation methods identify V. cholerae and make possible timely information regarding the genetic diversity of V. cholerae; our results set the stage for continued molecular epidemiological research to better understand the ... |
format |
Article in Journal/Newspaper |
author |
Amanda K Debes Jerome Ateudjieu Etienne Guenou Anna Lena Lopez Mark Philip Bugayong Pearl Joy Retiban Marcelino Garrine Inacio Mandomando Shan Li O Colin Stine David A Sack |
author_facet |
Amanda K Debes Jerome Ateudjieu Etienne Guenou Anna Lena Lopez Mark Philip Bugayong Pearl Joy Retiban Marcelino Garrine Inacio Mandomando Shan Li O Colin Stine David A Sack |
author_sort |
Amanda K Debes |
title |
Evaluation in Cameroon of a Novel, Simplified Methodology to Assist Molecular Microbiological Analysis of V. cholerae in Resource-Limited Settings. |
title_short |
Evaluation in Cameroon of a Novel, Simplified Methodology to Assist Molecular Microbiological Analysis of V. cholerae in Resource-Limited Settings. |
title_full |
Evaluation in Cameroon of a Novel, Simplified Methodology to Assist Molecular Microbiological Analysis of V. cholerae in Resource-Limited Settings. |
title_fullStr |
Evaluation in Cameroon of a Novel, Simplified Methodology to Assist Molecular Microbiological Analysis of V. cholerae in Resource-Limited Settings. |
title_full_unstemmed |
Evaluation in Cameroon of a Novel, Simplified Methodology to Assist Molecular Microbiological Analysis of V. cholerae in Resource-Limited Settings. |
title_sort |
evaluation in cameroon of a novel, simplified methodology to assist molecular microbiological analysis of v. cholerae in resource-limited settings. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2016 |
url |
https://doi.org/10.1371/journal.pntd.0004307 https://doaj.org/article/ab92b3cd8dd24d39ae851ace1601b91c |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 10, Iss 1, p e0004307 (2016) |
op_relation |
http://europepmc.org/articles/PMC4703203?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0004307 https://doaj.org/article/ab92b3cd8dd24d39ae851ace1601b91c |
op_doi |
https://doi.org/10.1371/journal.pntd.0004307 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
10 |
container_issue |
1 |
container_start_page |
e0004307 |
_version_ |
1766347099126890496 |