Rapid and Accurate Diagnosis of the Respiratory Disease Pertussis on a Point-of-Care Biochip

Abstract

Background Pertussis is a highly contagious respiratory disease caused by the bacterium Bordetella pertussis ( B. pertussis ). The infection is difficult to diagnose especially in underserved or resource-limited areas. We developed a low-cost and instrument-free diagnostic method for rapid and accurate detection of B. pertussis on a point-of-care (POC) testing device. Methods We developed a paper/polymer hybrid microfluidic biochip integrated with loop-mediated isothermal amplification (LAMP) method for the rapid and accurate detection of B. pertussis . This microfluidic approach was validated by testing 100 de-identified remnant clinical nasopharyngeal swabs and aspirates, which were confirmed to be either positive or negative for B. pertussis by a validated real-time PCR assay at the Children's Hospital Los Angeles. Findings The instrument-free detection results could be successfully read by the naked eye within 45 min with a limit of detection (LOD) of 5 DNA copies per well. Our optimized bacterial lysis protocol allowed the direct testing of clinical samples without any complicated sample processing/preparation (i.e. DNA extraction) or the use of any equipment (e.g. centrifuges). The validation of the microfluidic approach was accomplished by testing 100 clinical samples. High sensitivity (100%) and specificity (96%) with respect to real-time PCR were achieved. Interpretation This microfluidic biochip shows great potential for point-of-care disease diagnosis in various venues including schools and physician's offices, especially in low-resource settings in developing nations. Funding NIH/NIAID under award number R21AI107415, NIH RCMI Pilot Grant, the Philadelphia Foundation, the Medical Center of the Americas Foundation.

Advances in point-of-care nucleic acid extraction technologies for rapid diagnosis of human and plant diseases

“…The detection sensitivity of the LAMP assay using boiled samples was approximately 90%. Dou et al (2019) directly added cell lysate to the LAMP mixture to detect Bordetella pertussis from nasopharyngeal swab samples. For cell lysis, an equal volume of sample and bacterial lysis buffer (50 mM Tris buffer, 4 M urea, and 0.1% Triton, pH 7.5) were mixed and incubated at room temperature for 10 min.…”

Immunomodulation as a Novel Strategy for Prevention and Treatment of Bordetella spp. Infections

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A fully automated centrifugal microfluidic system for sample-to-answer viral nucleic acid testing

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