期刊
ECLINICALMEDICINE
卷 8, 期 -, 页码 72-77出版社
ELSEVIER
DOI: 10.1016/j.eclinm.2019.02.008
关键词
Pertussis diagnosis; Point-of-care detection; Microfluidic biochip; Whooping cough; Loop-mediated isothermal amplification (LAMP)
资金
- National Institute of Allergy and Infectious Disease of the NIH [R21AI107415]
- Philadelphia Foundation
- Medical Center of the Americas Foundation
- U.S. NSF-PREM program [DMR 1205302, 1827745]
- National Institute of General Medical Sciences of the NIH [SC2GM105584]
- NIH RCMI Pilot grant
- NIH BUILDing Scholar Summer Sabbatical Award (NIGMS) [RL5GM118969, TL4GM118971, UL1GM11897]
- University of Texas at El Paso (UTEP)
- University of Texas System
- National Institutes on Minority Health and Health Disparities (NIMHD) [G12MD007592]
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 5DNA copies perwell. 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. (C) 2019 Published by Elsevier Ltd.
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