A comparative analysis of preservation techniques for the optimal molecular detection of hookworm DNA in a human fecal specimen.
Proper collection and storage of fecal samples is necessary to guarantee the subsequent reliability of DNA-based soil-transmitted helminth diagnostic procedures. Previous research has examined various methods to preserve fecal samples for subsequent microscopic analysis or for subsequent determinati...
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ftdoajarticles:oai:doaj.org/article:c9c0bb1ef91f40398bb683cbe4634e21 2023-05-15T15:15:44+02:00 A comparative analysis of preservation techniques for the optimal molecular detection of hookworm DNA in a human fecal specimen. Marina Papaiakovou Nils Pilotte Ben Baumer Jessica Grant Kristjana Asbjornsdottir Fabian Schaer Yan Hu Raffi Aroian Judd Walson Steven A Williams 2018-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0006130 https://doaj.org/article/c9c0bb1ef91f40398bb683cbe4634e21 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC5773002?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0006130 https://doaj.org/article/c9c0bb1ef91f40398bb683cbe4634e21 PLoS Neglected Tropical Diseases, Vol 12, Iss 1, p e0006130 (2018) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2018 ftdoajarticles https://doi.org/10.1371/journal.pntd.0006130 2022-12-31T15:00:28Z Proper collection and storage of fecal samples is necessary to guarantee the subsequent reliability of DNA-based soil-transmitted helminth diagnostic procedures. Previous research has examined various methods to preserve fecal samples for subsequent microscopic analysis or for subsequent determination of overall DNA yields obtained following DNA extraction. However, only limited research has focused on the preservation of soil-transmitted helminth DNA in stool samples stored at ambient temperature or maintained in a cold chain for extended periods of time.Quantitative real-time PCR was used in this study as a measure of the effectiveness of seven commercially available products to preserve hookworm DNA over time and at different temperatures. Results were compared against "no preservative" controls and the "gold standard" of rapidly freezing samples at -20°C. The preservation methods were compared at both 4°C and at simulated tropical ambient temperature (32°C) over a period of 60 days. Evaluation of the effectiveness of each preservative was based on quantitative real-time PCR detection of target hookworm DNA.At 4°C there were no significant differences in DNA amplification efficiency (as measured by Cq values) regardless of the preservation method utilized over the 60-day period. At 32°C, preservation with FTA cards, potassium dichromate, and a silica bead two-step desiccation process proved most advantageous for minimizing Cq value increases, while RNA later, 95% ethanol and Paxgene also demonstrate some protective effect. These results suggest that fecal samples spiked with known concentrations of hookworm-derived egg material can remain at 4°C for 60 days in the absence of preservative, without significant degradation of the DNA target. Likewise, a variety of preservation methods can provide a measure of protection in the absence of a cold chain. As a result, other factors, such as preservative toxicity, inhibitor resistance, preservative cost, shipping requirements, sample infectivity, and labor costs ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 12 1 e0006130 |
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 Marina Papaiakovou Nils Pilotte Ben Baumer Jessica Grant Kristjana Asbjornsdottir Fabian Schaer Yan Hu Raffi Aroian Judd Walson Steven A Williams A comparative analysis of preservation techniques for the optimal molecular detection of hookworm DNA in a human fecal specimen. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Proper collection and storage of fecal samples is necessary to guarantee the subsequent reliability of DNA-based soil-transmitted helminth diagnostic procedures. Previous research has examined various methods to preserve fecal samples for subsequent microscopic analysis or for subsequent determination of overall DNA yields obtained following DNA extraction. However, only limited research has focused on the preservation of soil-transmitted helminth DNA in stool samples stored at ambient temperature or maintained in a cold chain for extended periods of time.Quantitative real-time PCR was used in this study as a measure of the effectiveness of seven commercially available products to preserve hookworm DNA over time and at different temperatures. Results were compared against "no preservative" controls and the "gold standard" of rapidly freezing samples at -20°C. The preservation methods were compared at both 4°C and at simulated tropical ambient temperature (32°C) over a period of 60 days. Evaluation of the effectiveness of each preservative was based on quantitative real-time PCR detection of target hookworm DNA.At 4°C there were no significant differences in DNA amplification efficiency (as measured by Cq values) regardless of the preservation method utilized over the 60-day period. At 32°C, preservation with FTA cards, potassium dichromate, and a silica bead two-step desiccation process proved most advantageous for minimizing Cq value increases, while RNA later, 95% ethanol and Paxgene also demonstrate some protective effect. These results suggest that fecal samples spiked with known concentrations of hookworm-derived egg material can remain at 4°C for 60 days in the absence of preservative, without significant degradation of the DNA target. Likewise, a variety of preservation methods can provide a measure of protection in the absence of a cold chain. As a result, other factors, such as preservative toxicity, inhibitor resistance, preservative cost, shipping requirements, sample infectivity, and labor costs ... |
format |
Article in Journal/Newspaper |
author |
Marina Papaiakovou Nils Pilotte Ben Baumer Jessica Grant Kristjana Asbjornsdottir Fabian Schaer Yan Hu Raffi Aroian Judd Walson Steven A Williams |
author_facet |
Marina Papaiakovou Nils Pilotte Ben Baumer Jessica Grant Kristjana Asbjornsdottir Fabian Schaer Yan Hu Raffi Aroian Judd Walson Steven A Williams |
author_sort |
Marina Papaiakovou |
title |
A comparative analysis of preservation techniques for the optimal molecular detection of hookworm DNA in a human fecal specimen. |
title_short |
A comparative analysis of preservation techniques for the optimal molecular detection of hookworm DNA in a human fecal specimen. |
title_full |
A comparative analysis of preservation techniques for the optimal molecular detection of hookworm DNA in a human fecal specimen. |
title_fullStr |
A comparative analysis of preservation techniques for the optimal molecular detection of hookworm DNA in a human fecal specimen. |
title_full_unstemmed |
A comparative analysis of preservation techniques for the optimal molecular detection of hookworm DNA in a human fecal specimen. |
title_sort |
comparative analysis of preservation techniques for the optimal molecular detection of hookworm dna in a human fecal specimen. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2018 |
url |
https://doi.org/10.1371/journal.pntd.0006130 https://doaj.org/article/c9c0bb1ef91f40398bb683cbe4634e21 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 12, Iss 1, p e0006130 (2018) |
op_relation |
http://europepmc.org/articles/PMC5773002?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0006130 https://doaj.org/article/c9c0bb1ef91f40398bb683cbe4634e21 |
op_doi |
https://doi.org/10.1371/journal.pntd.0006130 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
12 |
container_issue |
1 |
container_start_page |
e0006130 |
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1766346081089617920 |