Evidence map›Paper›PMID 39898969›Full record

ArticleACS synthetic biology2025

Harnessing Non-standard Nucleic Acids for Highly Sensitive Icosaplex (20-Plex) Detection of Microbial Threats for Environmental Surveillance.

Hinako Kawabe, Luran Manfio, Sebastian Magana Pena, Nicolette A Zhou, Kevin M Bradley, Cen Chen, Chris McLendon, Steven A Benner, Karen Levy, Zunyi Yang and 2 more

Abstract read
In one paragraph

Article in ACS synthetic biology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers, 1 of them a synthesis that pooled it.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed, 1 pooled it
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

4 citing papers in PubMed, 1 synthesis or guideline pooled it.

  1. Pooled it
  2. Article
  3. Article
  4. Article
4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

12 authors.

Hinako KawabeChemical Engineering, University of Washington, Seattle, Washington 98195, United States.
Luran ManfioFoundation for Applied Molecular Evolution (FfAME), 13709 Progress Blvd, Alachua, Florida 32615, United States.
Sebastian Magana PenaFoundation for Applied Molecular Evolution (FfAME), 13709 Progress Blvd, Alachua, Florida 32615, United States.
Nicolette A ZhouDepartment of Environmental and Occupational Health Sciences, University of Washington, Seattle, Washington 98195, United States.
Kevin M BradleyFoundation for Applied Molecular Evolution (FfAME), 13709 Progress Blvd, Alachua, Florida 32615, United States.
Cen ChenFoundation for Applied Molecular Evolution (FfAME), 13709 Progress Blvd, Alachua, Florida 32615, United States.ORCID 0000-0002-6883-9369
Chris McLendonFirebird Biomolecular Sciences LLC, 13709 Progress Blvd, Box 17, Alachua, Florida 32615, United States.
Steven A BennerFoundation for Applied Molecular Evolution (FfAME), 13709 Progress Blvd, Alachua, Florida 32615, United States.
Karen LevyDepartment of Environmental and Occupational Health Sciences, University of Washington, Seattle, Washington 98195, United States.ORCID 0000-0002-0968-9401
Zunyi YangFoundation for Applied Molecular Evolution (FfAME), 13709 Progress Blvd, Alachua, Florida 32615, United States.
Jorge A MarchandChemical Engineering, University of Washington, Seattle, Washington 98195, United States.ORCID 0000-0002-7765-610X
Erica R FuhrmeisterDepartment of Environmental and Occupational Health Sciences, University of Washington, Seattle, Washington 98195, United States.ORCID 0000-0001-5264-7533

Funding

XENOBIOTIC BIOTRANSFORMATION AND DISPOSITIONP30ES007033 · NIEHS · UNIVERSITY OF WASHINGTON · PI Nicole Ann Errett · 1995 to 2026
$42.5M
Gut microbiome, enteric infections and child growth across a rurual urban gradientR01AI137679 · NIAID · UNIVERSITY OF WASHINGTON · PI EISENBERG, JOSEPH N. S., LEVY, KAREN · 2018 to 2023
$5.3M
Basic Research for Diagnostics and Surveillance in Lower Resource EnvironmentsR01AI135146 · NIAID · FOUNDATION FOR APPLIED MOLECULAR EVOLUTN · PI BENNER, STEVEN A · 2021 to 2024
$2.8M
NIAID NIH HHS R01 AI135146NIAID NIH HHS R01 AI137679NIEHS NIH HHS P30 ES007033
6 · The paper itself

Abstract

Environmental surveillance and clinical diagnostics heavily rely on the polymerase chain reaction (PCR) for target detection. A growing list of microbial threats warrants new PCR-based detection methods that are highly sensitive, specific, and multiplexable. Here, we introduce a PCR-based icosaplex (20-plex) assay for detecting 18 enteropathogen and two antimicrobial resistance genes. This multiplexed PCR assay leverages the self-avoiding molecular recognition system (SAMRS) to avoid primer dimer formation, the artificially expanded genetic information system (AEGIS) for amplification specificity, and next-generation sequencing for amplicon identification. Using parallelized multitarget TaqMan Array Cards (TAC) to benchmark performance of the 20-plex assay on wastewater, soil, and human stool samples, we found 90% agreement on positive calls and 89% agreement on negative calls. Additionally, we show how long-read and short-read sequencing information from the 20-plex can be used to further classify allelic variants of genes and distinguish subspecies. The strategy presented offers sensitive, affordable, and robust multiplex detection that can be used to support efforts in wastewater-based epidemiology, environmental monitoring, and human/animal diagnostics.

Indexed as

Environmental MonitoringMultiplex Polymerase Chain ReactionFecesHigh-Throughput Nucleotide SequencingHumansWastewaterWastewaterAEGISenteric pathogensmultiplex PCRnext-generation sequencingSAMRSwastewater-based epidemiology

Identifiers

PMID39898969
PMCPMC11854376

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.