114: One-hour extraction-free LAMP HPV test for point-of-care screening

Barra MJ et al et al., Nature Communications (2025) 16:7295 - This study reports development and analytic evaluation of an extraction-free DARQ LAMP assay detecting HPV16, HPV18, and HPV45 plus a cellular control with a <1 hour sample-to-answer workflow on a low-cost benchtop heater/fluorimeter. Clinical testing in Houston (n=38) and Maputo (n=191) showed 100% and 93% concordance, respectively, with the GeneXpert reference test. Key terms: HPV16, HPV18, HPV45, LAMP, point-of-care.

Study Highlights:
The authors developed a multiplexed DARQ LAMP assay targeting HPV16, HPV18, HPV45 and a cellular control and implemented an extraction-free NaOH lysis allowing direct addition of lysate to the reaction. Limits of detection with extracted DNA were 57, 11.7, and 14.3 copies/reaction for HPV16, HPV18, and HPV45, respectively, and with crude lysate were ~30.4 SiHa, 11.7 HeLa, and <10 MS751 cells/reaction. The assay runs on a portable Axxin T8-ISO device in under one hour and achieved 100% concordance in Houston (n=38) and 93% concordance in Maputo (n=191) versus GeneXpert. Extraction-free workflow simplified specimen handling but contributed to reduced sensitivity for samples with low viral load.

Conclusion:
An extraction-free, multiplexed DARQ LAMP assay provides rapid (<1 h), sensitive detection of HPV16/18/45 with high concordance to a WHO-prequalified reference test and a simple workflow suitable for point-of-care use in resource-limited settings, while further clinical validation and expansion to additional HPV types are needed.

Music:
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Article title:
One-hour extraction-free loop-mediated isothermal amplification HPV DNA assay for point-of-care testing in Maputo, Mozambique

First author:
Barra MJ et al

Journal:
Nature Communications (2025) 16:7295

DOI:
10.1038/s41467-025-62454-x

Reference:
Barra MJ et al. One-hour extraction-free loop-mediated isothermal amplification HPV DNA assay for point-of-care testing in Maputo, Mozambique. Nature Communications (2025) 16:7295. https://doi.org/10.1038/s41467-025-62454-x

License:
This episode is based on an open-access article published under the Creative Commons Attribution 4.0 International License (CC BY 4.0) – https://creativecommons.org/licenses/by/4.0/

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Episode link: https://basebybase.com/episodes/one-hour-extraction-free-loop-mediated-isothermal-amplification-hpv-dna-assay-for-point-of-care-testing-in-maputo-mozambique

QC:
This episode was checked against the original article PDF and publication metadata for the episode release published on 2025-08-22.

QC Scope:
- article metadata and core scientific claims from the narration
- excludes analogies, intro/outro, and music
- transcript coverage: Audited the transcript’s depiction of the DARQ LAMP extraction-free HPV assay, its workflow, device, clinical evaluations, and performance metrics.
- transcript topics: HPV biology and cervical cancer screening context; DARQ LAMP methodology and extraction-free lysate; Targets HPV16/HPV18/HPV45 and cellular control; Point-of-care device and workflow (65 C, T8-ISO); Clinical evaluation in Houston and Maputo; Performance metrics, concordance, and limitations

QC Summary:
- factual score: 10/10
- metadata score: 10/10
- supported core claims: 8
- claims flagged for review: 0
- metadata checks passed: 4
- metadata issues found: 0

Metadata Audited:
- article_doi
- article_title
- article_journal
- license

Factual Items Audited:
- Targets HPV16/18/45 and cellular gDNA control
- Extraction-free lysate prep using NaOH
- Four-step protocol on the T8-ISO yielding results in under 1 hour
- Clinical concordance: Houston 100% (n=38) and Maputo 93% (n=191)
- LOD with extracted DNA: HPV16 57 copies/reaction; HPV18 11.7 copies/reaction; HPV45 14.3 copies/reaction
- LOD with crude lysate: HPV16 30.4 SiHa cells; HPV18 11.7 HeLa cells; HPV45 <10 MS751 cells

QC result: Pass.