A Novel, Multi-Parallel, Real-Time Polymerase Chain Reaction Approach for Eight Gastrointestinal Parasites Provides Improved Diagnostic Capabilities to Resource-Limited At-Risk Populations
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Diagnosis of gastrointestinal parasites has traditionally relied on stool microscopy, which has low diagnostic sensitivity and specificity. We have developed a novel, rapid, high-throughput quantitative multi-parallel real-time polymerase chain reaction (qPCR) platform. Species-specific primers/probes were used for eight common gastrointestinal parasite pathogens: Ascaris lumbricoides, Necator americanus, Ancylostoma duodenale, Giardia lamblia, Cryptosporidium spp., Entamoeba histolytica, Trichuris trichiura, and Strongyloides stercoralis. Stool samples from 400 13-month-old children in rural Ecuador were analyzed and the qPCR was compared with a standard direct wet mount slide for stool microscopy, as were 125 8–14-year-old children before and after anthelmintic treatment. The qPCR showed higher detection rates for all parasites compared with direct microscopy, Ascaris (7.0% versus 5.5%) and for Giardia (31.5% versus 5.8%). Using an enhanced DNA extraction method, we were able to detect T. trichiura DNA. These assays will be useful to refine treatment options for affected populations, ultimately leading to better health outcomes.
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Primers, probes, and protocols for the qPCR, including methods for generating a recombinant plasmid standard curve, were taken from Mejia et al. (2013).
“Primers, probes, and protocols for the qPCR, including methods for generating a recombinant plasmid containing target E. histolytica sequence to make a standard curve, were taken from Mejia et al. (2013) [95].”
From Population structure of human gut bacteria in a diverse cohort from rural Tanzania and Botswana (2019) · cites this paper as [95]
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- Authors
- Mejia, R., Vicuña, Y., Broncano, N., Sandoval, C., Vaca, M., Chico, M., Cooper, P. J., & Nutman, T. B.
- Corresponding author
- Nutman, Thomas B.
- Venue
- The American Society of Tropical Medicine and Hygiene
- Published
- 2013-06-05
- Volume
- 88
- Issue
- 6
- Pages
- 1041-1047
- ISSN
- 0002-9637 (ISSN-L)
- 1476-1645
- PMC
- PMC3752800
- OpenAlex
- W2123559199
- Cited by
- 273
- References
- 26
- Citations by year
- 2026: 12 · 2025: 17 · 2024: 24 · 2023: 15 · 2022: 19 · 2021: 29 · 2020: 35 · 2019: 19 · 2018: 23 · 2017: 25 · 2016: 19 · 2015: 20 · 2014: 12 · 2013: 4
- FWCI
- 15.45
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- American Journal of Tropical Medicine and Hygiene (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- LSTM Online Archive (Liverpool School of Tropical Medicine) (submittedVersion)
- Funders
- Wellcome Trust (088862)
- Topics
- Life Sciences › Immunology and Microbiology › Parasitology › Parasites and Host Interactions
- Life Sciences › Immunology and Microbiology › Parasitology › Parasitic Infections and Diagnostics
- Health Sciences › Veterinary › Small Animals › Helminth infection and control
- Keywords
- gastrointestinal parasites, parasitological diagnosis, stool microscopy, Ascaris lumbricoides, Necator americanus, Ancylostoma duodenale, Giardia lamblia, Cryptosporidium, Entamoeba histolytica, Trichuris trichiura, Strongyloides stercoralis, anthelmintic treatment, children, resource-limited settings, DNA extraction
- MeSH terms
- Adolescent, Ancylostoma, Animals, Anthelmintics, Child, Cryptosporidium, Ecuador, Entamoeba histolytica, Feces, Humans, India, Infant, Intestinal Diseases, Parasitic, Mali, Microscopy, North America, Rural Population, Sensitivity and Specificity, Species Specificity, Trichuris, Reproducibility of Results, Prevalence, Giardia lamblia, Ascaris lumbricoides, Strongyloides stercoralis, Necator americanus, DNA Primers, DNA, Helminth, Empirical Research, High-Throughput Screening Assays, Real-Time Polymerase Chain Reaction
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