Loading...
Comparison of multi-parallel quantitative real-time PCRs targeting different DNA regions and detecting soil-transmitted helminths in stool
Citations
Altmetric:
Advisors
Editors
Other Contributors
Affiliation
EPub Date
Issue Date
2024-09
Submitted Date
Subject Terms
Collections
Files
Loading...
Published version
Adobe PDF, 803.14 KB
Research Projects
Organizational Units
Journal Issue
Other Titles
Abstract
<jats:title>Abstract</jats:title><jats:sec>
<jats:title>Background</jats:title>
<jats:p>Soil-transmitted helminths infect an estimated 18% of the world’s population, causing a significant health burden. Microscopy has been the primary tool for diagnosing eggs from fecal samples, but its sensitivity drops in low-prevalence settings. Quantitative real-time polymerase chain reaction (qPCR) is slowly increasing in research and clinical settings. However, there is still no consensus on preferred qPCR targets.</jats:p>
</jats:sec><jats:sec>
<jats:title>Methods</jats:title>
<jats:p>We aimed to compare soil-transmitted helminth (STH) DNA detection methods by testing naïve stool samples spiked with known quantities of STH eggs and larvae. DNA extracts from spiked samples were tested using independent quantitative realtime PCR (qPCR) assays targeting ribosomal or putative non-protein coding satellite sequences.</jats:p>
</jats:sec><jats:sec>
<jats:title>Results</jats:title>
<jats:p>For <jats:italic>Trichuris trichiura,</jats:italic> there was a strong correlation between egg/larvae counts and qPCR results using either qPCR method (0.86 and 0.87, respectively). Strong correlations also existed for <jats:italic>A. lumbricoides</jats:italic> (0.60 and 0.63, respectively), but weaker correlations were found for <jats:italic>Ancylostoma duodenale</jats:italic> (0.41 for both assays) and <jats:italic>Strongyloides stercoralis</jats:italic> (0.48 and 0.65, respectively). No correlation for <jats:italic>Necator americanus</jats:italic> was observed when testing with either qPCR assay. Both assays had fair-to-moderate agreement across targets when using field-collected stool samples (0.28–0.45, for all STHs), except for <jats:italic>S. stercoralis</jats:italic> (0.12) with slight agreement.</jats:p>
</jats:sec><jats:sec>
<jats:title>Conclusions</jats:title>
<jats:p>There is a strong correlation between qPCR results and egg/larvae counts. Our study confirms that qPCR is an effective diagnostic tool, even with low-intensity infections, regardless of the DNA-based diagnostic marker used. However, the moderate agreement between the two different qPCR assays when testing field samples highlights the need to understand the role of these targets in the genome so that the parasite burden can be quantified more accurately and consistently by qPCR.</jats:p>
</jats:sec><jats:sec>
<jats:title>Graphical abstract</jats:title>
</jats:sec>
Citation
Publisher
Journal
Research Unit
PubMed ID
PubMed Central ID
Embedded videos
Type
Journal Article
Item Description
Series/Report no.
ISSN
1756-3305
EISSN
1756-3305