Evaluation in Cameroon of a Novel, Simplified Methodology to Assist Molecular Microbiological Analysis of V. cholerae in Resource-Limited Settings

Debes, Amanda K.; Ateudjieu, Jérôme; Guenou, Etienne; Lopez, Anna Lena; Bugayong, Mark Philip; Retiban, Pearl Joy; Garrine, Marcelino; Mandomando, Inácio; Li, Shan; Stine, O. Colin; Sack, David A.

doi:10.1371/journal.pntd.0004307

Cited by 19 publications

(28 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The Chelex method and similar boiling methods have been previously validated for the extraction of DNA from samples containing bacterial pathogens preserved on Whatman cards or filter paper. 21,25 In these previous studies, diarrheal stool or water samples were analyzed. However, to our knowledge, our study is the first to investigate this method for parasites on non-diarrheal stool.…”

Section: Discussionmentioning

confidence: 99%

“…Previous work in Cameroon has shown that fecal specimens can be preserved on Whatman 903 protein saver cards for subsequent DNA extraction and PCR analysis for Vibrio cholerae. 21 Furthermore, another study has demonstrated that Whatman FTA Elute cards (Sigma Aldrich, St. Louis, MO) can be used to preserve fecal samples containing bacterial and parasitic enteropathogens for PCR detection. 22 Whatman 903 protein saver cards have the advantage of being a third of the cost of Whatman FTA Elute cards (1.5 USD versus 4.60 USD per card), which will allow for a more economically feasible option in low-resource settings.…”

Section: Introductionmentioning

confidence: 99%

“…To date the method of storing stool samples on Whatman 903 protein saver cards has only been validated for Vibrio cholerae. 21 Research is needed on the efficacy of this method for other enteric pathogens.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Whatman Protein Saver Cards for Storage and Detection of Parasitic Enteropathogens

Natarajan

Kabir

Perin

et al. 2018

The American Journal of Tropical Medicine and Hygiene

View full text Add to dashboard Cite

Current methods to identify the etiology of diarrhea require laboratory facilities for storage of pathogens, which is often challenging in low-resource settings. This study evaluated the efficacy of a low-cost method for preserving stool specimens for the detection of parasitic enteropathogens using Whatman 903 protein saver cards (Sigma-Aldrich, St. Louis, MO). Stool samples known to be positive by multiplex real-time polymerase chain reaction for Giardia lamblia, Cryptosporidium spp., and Entamoeba histolytica parasites were preserved on 232 Whatman cards. DNA was then extracted from cards using Chelex and Qiagen extraction protocols, and tested for these parasites using multiplex realtime PCR. We included stool samples known to have a higher parasite load (cycle threshold [ct]-value < 30) and those with a lower parasite load (ct values 30-35). Sensitivities and specificities were determined using DNA extracted directly from whole stool samples using Qiagen kits (QIAGEN, Hilden, Germany). For whole stool samples with ct values < 30, preserved directly on Whatman 903 protein saver cards for Giardia analysis, the sensitivity was 100% for both Qiagen and Chelex DNA extraction. For E. histolytica, this was 100% for sensitivity for Qiagen and 80% for Chelex DNA extractions, and for Cryptosporidium, this was 80% for Qiagen and 50% for Chelex DNA extraction. The specificity was 100% for all parasites for all extraction procedures. Given the high sensitivity for stool samples with higher parasite loads, we recommend the use of the Whatman 903 protein saver card for preserving fecal specimens for the analysis of Giardia and E. histolytica using Qiagen DNA extractions in low-resource settings.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Whatman Protein Saver Cards for Storage and Detection of Parasitic Enteropathogens

Natarajan

Kabir

Perin

et al. 2018

The American Journal of Tropical Medicine and Hygiene

View full text Add to dashboard Cite

show abstract

“…The audience learned from Martin Mengel, from Agence Medicine Preventive, Paris, that further efforts to understand the distribution of cholera in Africa have been undertaken by AfriChol, the African Cholera Surveillance Network (19). To further understand disease transmission patterns, Amanda Debes from the JHU Bloomberg School of Public Health described a combined clinical and environmental surveillance of Vibrio cholerae for resourcelimited areas that involves testing of clinical specimens with a rapid dipstick assay (crystal V. cholerae; Span Diagnostics Ltd., India), after alkaline peptone water (APW) enrichment, and blotting onto filter paper for subsequent analysis using molecular methods (20,21). This method was successfully used in the far north region of Cameroon and enabled early detection of cases and rapid response (21).…”

mentioning

confidence: 99%

Highlights of the 8th International Conference on Vaccines for Enteric Diseases: the Scottish Encounter To Defeat Diarrheal Diseases

Tennant

Steele

Pasetti

2016

Clin Vaccine Immunol

View full text Add to dashboard Cite

Infectious diarrhea is a leading cause of morbidity and of mortality; the burden of disease affects individuals of all ages but particularly young children, especially those living in poor regions where the disease is endemic. It is also a health concern for international travelers to these areas. Experts on vaccines and enteric infections and advocates for global health improvement gathered in Scotland from 8 to 10 July 2015 to discuss recent advances in the assessment and understanding of the burden of enteric diseases and progress in the development and implementation of strategies to prevent these infections. Highlights of the meeting included description of advances in molecular assays to estimate pathogen-specific prevalence, methods to model epidemiologic trends, novel approaches to generate broad-spectrum vaccines, new initiatives to evaluate vaccine performance where they are most needed, renewed interest in human challenge models, immunological readouts as predictors of vaccine efficacy, maternal immunization to prevent enteric infections, and the impact of maternal immunity on the vaccine take of infants. A follow-up scientific gathering to advance Shigella and enterotoxigenic Escherichia coli (ETEC) vaccine efforts will be held from 28 to 30 June 2016 in Washington, DC. E nteric infections and diarrhea represent a major global cause of disease burden and of death and disability, primarily in lessdeveloped regions of the world (1). The most affected are children under 5 years of age and special groups, e.g., immunocompromised, the elderly, and pregnant women. Acute infectious diarrhea is also a health concern for travelers to areas where the disease is endemic, as it may require hospitalization and have chronic sequelae. Scientists from academia and government, vaccine producers, and public health advocates, including funding sponsors from around the world, gathered at the 8th International Conference on Vaccines for Enteric Diseases (VED) in Edinburgh, Scotland, to discuss recent progress in our understanding of enteric disease etiology, new methods of surveillance, the increasing awareness of the long-lasting health impairment associated with enteric infections, the development and evaluation of new vaccine candidates, and introduction of vaccines into existing immunization schedules and in emergency situations and in high-risk groups, among many other topics. Knowledge gaps were identified, as was the need for more frequent exchange of information and interactions within the research community to accelerate the development and implementation of preventive tools. This meeting highlight summarizes the main findings presented at the VED meeting and discussions and recommendations made, particularly the progress in vaccine development and clinical evaluation efforts.Prevalence of Shigella and ETEC and broad effects associated with infection. The Global Enteric Multicenter Study (GEMS) revealed that Shigella and enterotoxigenic Escherichia coli (ETEC) are among the top five major causes of moderate to s...

show abstract

“…The authors described the novel use of filter paper to preserve DNA specimens for polymerase chain reaction (PCR) confirmation of cholera. In another recent article, 2 they used the same sampling method to genotype Vibrio cholerae using multiple loci variable number tandem repeat (VNTR) analysis (MLVA), in the same resource-constrained area. In both studies, stool specimens were initially enriched for 6–8 hours in alkaline peptone water (APW).…”

mentioning

confidence: 99%

Direct Dried Stool Sampling on Filter Paper for Molecular Analyses of Cholera

Moore

Normand

Koivogui

et al. 2016

The American Society of Tropical Medicine and Hygiene

View full text Add to dashboard Cite

Evaluation in Cameroon of a Novel, Simplified Methodology to Assist Molecular Microbiological Analysis of V. cholerae in Resource-Limited Settings

Cited by 19 publications

References 22 publications

Whatman Protein Saver Cards for Storage and Detection of Parasitic Enteropathogens

Whatman Protein Saver Cards for Storage and Detection of Parasitic Enteropathogens

Highlights of the 8th International Conference on Vaccines for Enteric Diseases: the Scottish Encounter To Defeat Diarrheal Diseases

Direct Dried Stool Sampling on Filter Paper for Molecular Analyses of Cholera

Contact Info

Product

Resources

About