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<p>Canine blood parasites, including Hepatozoon canis, Babesia canis, Babesia gibsoni, Ehrlichia canis, and Anaplasma platys, pose significant health risks to dogs and potential zoonotic threats, particularly in resource-limited settings where tick-borne diseases affect millions annually by Manzini et al (Clin Microbiol Rev 23:235–251, 2010). Traditional diagnostics like PCR require sophisticated equipment and expertise, delaying timely intervention. This study presents a fully integrated centrifugal microfluidic chip (IACMC) combined with a controlled-release reagent cartridge for point-of-care (POC) multiplex detection of these five parasites plus an internal control in spiked canine blood samples (<i>n</i> = 9 replicates). The chip, fabricated from injection-molded PMMA and sealed with pressure-sensitive adhesive (PSA), integrates a silica membrane for nucleic acid extraction, a cartridge with laser-activated wax-sealed reagent compartments (washing buffer 1, washing buffer 2, elution buffer, and diluent), and six detection wells pre-loaded with LAMP lyophilized microspheres. Optimized experiments achieved a detection limit of 100–1000 copies/mL, comparable to off-chip LAMP, with 100% sensitivity and specificity in tested spiked samples (95% CI: 70–100%; no cross-reactivity observed), delivering results in &lt; 45&#xa0;min. The portable detection device, featuring centrifugal, heating, laser, and fluorescence modules, automates the process, ensuring a closed, contamination-free workflow. This platform offers promising advancements for field-deployable veterinary diagnostics at a materials-based estimated cost of &lt;$3/test.</p>

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Fully integrated centrifugal microfluidic chip for point-of-care multiplex detection of canine blood parasites

  • 期刊论文

  • 发表日期：

    2026年1月14日

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  • Peijun He,
  • Haiqin Tan,
  • Tianhong Shen,
  • Chirenjeevi Krishnan,
  • Lin Li,
  • Hejin Wu,
  • Lin Chen,
  • Bo Yu,
  • Zhinan Xu

摘要

Canine blood parasites, including Hepatozoon canis, Babesia canis, Babesia gibsoni, Ehrlichia canis, and Anaplasma platys, pose significant health risks to dogs and potential zoonotic threats, particularly in resource-limited settings where tick-borne diseases affect millions annually by Manzini et al (Clin Microbiol Rev 23:235–251, 2010). Traditional diagnostics like PCR require sophisticated equipment and expertise, delaying timely intervention. This study presents a fully integrated centrifugal microfluidic chip (IACMC) combined with a controlled-release reagent cartridge for point-of-care (POC) multiplex detection of these five parasites plus an internal control in spiked canine blood samples (n = 9 replicates). The chip, fabricated from injection-molded PMMA and sealed with pressure-sensitive adhesive (PSA), integrates a silica membrane for nucleic acid extraction, a cartridge with laser-activated wax-sealed reagent compartments (washing buffer 1, washing buffer 2, elution buffer, and diluent), and six detection wells pre-loaded with LAMP lyophilized microspheres. Optimized experiments achieved a detection limit of 100–1000 copies/mL, comparable to off-chip LAMP, with 100% sensitivity and specificity in tested spiked samples (95% CI: 70–100%; no cross-reactivity observed), delivering results in < 45 min. The portable detection device, featuring centrifugal, heating, laser, and fluorescence modules, automates the process, ensuring a closed, contamination-free workflow. This platform offers promising advancements for field-deployable veterinary diagnostics at a materials-based estimated cost of <$3/test.