ArticlePLoS neglected tropical diseases2019
Comparison of four DNA extraction and three preservation protocols for the molecular detection and quantification of soil-transmitted helminths in stool.
Article in PLoS neglected tropical diseases, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 46 papers, 2 of them syntheses that pooled it.
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Who cites it
46 citing papers in PubMed, 2 syntheses or guidelines pooled it, 78 citations in OpenAlex.
- Transforming gastrointestinal helminth parasite identification in vertebrate hosts with metabarcoding: a systematic review.Parasites & vectors · 2024Pooled it
- Anthelmintic drugs for treating ascariasis.The Cochrane database of systematic reviews · 2020Pooled it
- Comparison of real-time PCR and the Kato-Katz method for the diagnosis of soil-transmitted helminthiasis and assessment of cure in a randomized controlled trial.BMC microbiology · 2020Trial
- Optimisation and analytical assessment of a TaqMan™ probe-based real-time PCR assay designed to diagnose infection with Schistosoma japonicum.Parasites & vectors · 2026Article
- Scoop That Poop: Optimising Faecal Sample Pre-Processing for Parasite Metabarcoding.Molecular ecology resources · 2026Article
- Widespread Trichuris incognita reveals hidden diversity and reshapes understanding of human whipworm infections.Nature communications · 2025Article
- Performance evaluation of the diaxxoPCR system for rapid and user-friendly stool-based diagnosis of trichuriasis, ascariasis and strongyloidiasis in Mozambique.PLoS neglected tropical diseases · 2025Article
- Development of loop mediated isothermal amplification assay for rapid detection and its comparison with PCR techniques for Ancylostoma duodenale.Scientific reports · 2025Article
- Article
- A survey of morphological, molecular, and histopathological characteristics ofFrontiers in veterinary science · 2025Article
- Molecular epidemiology and population genetics of Schistosoma mansoni infecting school-aged children situated along the southern shoreline of Lake Malawi, Malawi.PLoS neglected tropical diseases · 2024Article
- Revision of the GenusInsects · 2024Article
- Estimating Prevalence and Infection Intensity of Soil-Transmitted Helminths Using Quantitative Polymerase Chain Reaction and Kato-Katz in School-Age Children in Angola.The American journal of tropical medicine and hygiene · 2024Article
- Article
- Assemblages and Subassemblages ofJournal of parasitology research · 2024Article
- Molecular Epidemiology and Assemblage Typing of Giardia duodenalis in School-Age Children Situated along the Southern Shoreline of Lake Malawi, Malawi.The American journal of tropical medicine and hygiene · 2023Article
- Design of a Protocol for Soil-Transmitted Helminths (in Light of the NematodeDiagnostics (Basel, Switzerland) · 2023Article
- Optimizing DNA Extraction from Pediatric Stool for Diagnosis of Tuberculosis and Use in Next-Generation Sequencing Applications.Microbiology spectrum · 2023Article
- A Pilot Comparison of Fixatives for Hookworm Real-time Polymerase Chain Reaction.The American journal of tropical medicine and hygiene · 2023Article
- Faecal metabarcoding provides improved detection and taxonomic resolution for non-invasive monitoring of gastrointestinal nematode parasites in wild moose populations.Parasites & vectors · 2023Article
Corrections and comments
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Authors and funding
10 authors at 4 institutions in 4 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundA DNA extraction and preservation protocol that yields sufficient and qualitative DNA is pivotal for the success of any nucleic acid amplification test (NAAT), but it still poses a challenge for soil-transmitted helminths (STHs), including Ascaris lumbricoides, Trichuris trichiura and the two hookworms (Necator americanus and Ancylostoma duodenale). In the present study, we assessed the impact of different DNA extraction and preservativation protocols on STH-specific DNA amplification from stool. METHODOLOGY AND PRINCIPAL
findingsIn a first experiment, DNA was extracted from 37 stool samples with variable egg counts for T. trichiura and N. americanus applying two commercial kits, both with and without a prior bead beating step. The DNA concentration of T. trichiura and N. americanus was estimated by means of qPCR. The results showed clear differences in DNA concentration across both DNA extraction kits, which varied across both STHs. They also indicated that adding a bead beating step substantially improved DNA recovery, particularly when the FECs were high. In a second experiment, 20 stool samples with variable egg counts for A. lumbricoides, T. trichiura and N. americanus were preserved in either 96% ethanol, 5% potassium dichromate or RNAlater and were stored at 4°C for 65, 245 and 425 days. DNA was extracted using the DNeasy Blood & Tissue kit with a bead beating step. Stool samples preserved in ethanol proved to yield higher DNA concentrations as FEC increased, although stool samples appeared to be stable over time in all preservatives.
conclusionsThe choice of DNA extraction kit significantly affects the outcome of NAATs. Given the clear benefit of bead beating and our validation of ethanol for (long-term) preservation, we recommend that these aspects of the protocol should be adopted by any stool sampling and DNA extraction protocol for downstream NAAT-based detection and quantification of STHs.
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