ArticleFrontiers in cellular and infection microbiology2020
High-Throughput Multiparallel Enteropathogen Detection via Nano-Liter qPCR.
Article in Frontiers in cellular and infection microbiology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- Molecular Methods to DetectmedRxiv : the preprint server for health sciences · 2026Article
- Comparative evaluation of Sm1-7-qPCR systems incorporated with Maxwell kit-based and NaOH-based DNA extraction methods for the detection of Schistosoma mansoni infection.Infectious diseases of poverty · 2026Article
- Phage predation, disease severity and pathogen genetic diversity in cholera patients.bioRxiv : the preprint server for biology · 2024Article
- Simultaneous detection and quantification of multiple pathogen targets in wastewater.medRxiv : the preprint server for health sciences · 2023Article
- High-throughput low-cost nl-qPCR for enteropathogen detection: A proof-of-concept among hospitalized patients in Bangladesh.PloS one · 2021Article
- Gold Standard Cholera Diagnostics Are Tarnished by Lytic Bacteriophage and Antibiotics.Journal of clinical microbiology · 2020Article
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Authors and funding
4 authors.
Funding
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Abstract
Quantitative molecular diagnostic methods can effectively detect pathogen-specific nucleic acid sequences, but costs associated with multi-pathogen panels hinder their widespread use in research trials. Nano-liter qPCR (nL-qPCR) is a miniaturized tool for quantification of multiple targets in large numbers of samples based on assay parallelization on a single chip, with potentially significant cost-savings due to rapid throughput and reduced reagent volumes. We evaluated a suite of novel and published assays to detect 17 enteric pathogens using a commercially available nL-qPCR technology. Amplification efficiencies ranged from 88 to 98% (mean 91%) and were reproducible across four operators at two separate facilities. When applied to fecal material, assays were sensitive and selective (99.8% of DNA amplified were genes from the target organism). Due to nanofluidic volumes, detection limits were 1-2 orders of magnitude less sensitive for nL-qPCR than an enteric TaqMan Array Card (TAC). However, higher detection limits do not hinder detection of diarrhea-causing pathogen concentrations. Compared to TAC, nL-qPCR displayed 99% (95% CI 0.98, 0.99) negative percent agreement and 62% (95% CI 0.59, 0.65) overall positive percent agreement for presence of pathogens across diarrheal and non-diarrheal fecal samples. Positive percent agreement was 89% among samples with concentrations above the nL-qPCR detection limits. nL-qPCR assays showed an underestimation bias of 0.34 log
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