Comparison of multi-parallel quantitative real-time PCRs targeting different DNA regions and detecting soil-transmitted helminths in stool.

Autor: Papaiakovou M; Department of Biological Sciences, Smith College, Northampton, MA, 01063, USA. mpapaiakovou@gmail.com.; Biodiversity & Health, Natural History Museum, Cromwell Road, London, SW7 5BD, UK. mpapaiakovou@gmail.com.; Department of Veterinary Medicine, University of Cambridge, Cambridge, CB3 0ES, UK. mpapaiakovou@gmail.com., Cimino RO; Instituto de Investigación de Enfermedades Tropicales (IIET), Universidad Nacional de Salta. Sede Regional Orán, Salta, Argentina.; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., Pilotte N; Department of Biological Sciences, Smith College, Northampton, MA, 01063, USA.; Department of Biology, Quinnipiac University, Hamden, CT, 06518, USA., Dunn J; Department of Infectious Disease and Epidemiology, Imperial College London, London, W2 1PG, UK., Littlewood DTJ; Biodiversity & Health, Natural History Museum, Cromwell Road, London, SW7 5BD, UK., Williams SA; Department of Biological Sciences, Smith College, Northampton, MA, 01063, USA., Krolewiecki AJ; Instituto de Investigación de Enfermedades Tropicales (IIET), Universidad Nacional de Salta. Sede Regional Orán, Salta, Argentina.; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., Mejia R; Instituto de Investigación de Enfermedades Tropicales (IIET), Universidad Nacional de Salta. Sede Regional Orán, Salta, Argentina. rmejia@bcm.edu.; Section of Tropical Medicine, Department of Pediatrics, National School of Tropical Medicine, Texas Children's Hospital and Baylor College of Medicine, Houston, TX, 77030, USA. rmejia@bcm.edu.
Jazyk: angličtina
Zdroj: Parasites & vectors [Parasit Vectors] 2024 Sep 13; Vol. 17 (1), pp. 390. Date of Electronic Publication: 2024 Sep 13.
DOI: 10.1186/s13071-024-06464-6
Abstrakt: Background: 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.
Methods: 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.
Results: For Trichuris trichiura, 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 A. lumbricoides (0.60 and 0.63, respectively), but weaker correlations were found for Ancylostoma duodenale (0.41 for both assays) and Strongyloides stercoralis (0.48 and 0.65, respectively). No correlation for Necator americanus 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 S. stercoralis (0.12) with slight agreement.
Conclusions: 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.
(© 2024. The Author(s).)
Databáze: MEDLINE
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