Establishment of a visual mixed-dye loop mediated isothermal amplification assay for detecting Plasmodium knowlesi.

Colorimetric detection of loop-mediated isothermal amplification (LAMP) amplicons is one of the easiest and most direct methods among all detection techniques. However, not all indicators exhibit a noticeable colour change for both positive and negative reactions. In this study, we present a novel and visualized dual colour LAMP assay for the detection of Plasmodium knowlesi. The assay utilizes two types of dyes (phenol red and hydroxynaphthol blue) to facilitate a clear and distinguishable colour change upon successful amplification. The colour changes generated by the current single dye LAMP is low contrast and sometimes not significant. Thus, we propose a new approach to enlarge the variation colour range as well as improve its suitability for naked-eye observation. The dual colour system enhances the visual readout of the assay, thereby reducing false positives and negatives. To evaluate the performance of the mixed-dye LAMP, we test it with 55 clinical samples. By using nested PCR as the reference method, dual-colour LAMP showed 100% sensitivity (95% confidence interval (CI): 91.9-100%) and 100% specificity (95% CI: 78.2-100%). This mixed-dye LAMP assay offers a rapid, cost-effective, and user-friendly diagnostic tool for P. knowlesi malaria, particularly suitable for resource-limited settings.

• Publication year

  2025

• Venue

  Acta Tropica

• Publication date

  2025-07-01

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1016/j.actatropica.2025.107752PMID 40695401
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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