Point-of-care microdevices for simultaneous detection of multiple pertussis-like respiratory diseases from Bordetella species

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ABSTRACT Pertussis, also known as whooping cough, caused by B. pertussis and B. parapertussis, is the only vaccine-preventable infection that remains endemic in the U.S. Since pertussis vaccine- induced immunity is not permanent, the immunized host is vulnerable to re-infection later in life. Worldwide, there are an estimated 24.1 million cases of pertussis, and about 160,700 deaths per year from pertussis in children, most of which are from developing nations. These two Bordetella species are highly contagious and cause similar clinical symptoms, making them difficult to diagnose. Although B. parapertussis infection manifests in milder symptoms to B. pertussis- derived infection, both infections can be deadly, especially to children, the elderly, and immunocompromised patients. Current diagnostic methods are either slow or require costly instruments. Low-cost point-of-care (POC) detection of these two whooping cough-causing pathogens is greatly needed to meet the unmet demand for rapid diagnosis of whooping cough in resource-limited settings. During our recent preliminary study, we, for the first time, developed a low-cost microfluidic technology integrated with the loop-mediated isothermal DNA amplification (LAMP) for rapid POC detection of B. pertussis. Herein, the goal of this proposal is to develop a new low-cost paper/polymer hybrid microfluidic POC device integrated with LAMP for rapid, specific, and sensitive diagnosis of two major whooping cough-causing Bordetella species (B. pertussis and B. parapertussis) that can be used in various venues such as physicians’ offices, schools, and other low-resource settings. Our central hypothesis is that the integration of a low- cost portable paper-based microfluidic technology with specific DNA testing assays can provide a fast, accurate, and rapid diagnosis in resource-poor settings (US patent # 10,875,024). To accomplish our goal, we have developed three specific aims: (1) Develop and optimize LAMP assays for simultaneous detection of B. pertussis and B. parapertussis; (2) Develop a low-cost paper/polymer hybrid microfluidic POC device for the simultaneous instrument-free detection of B. pertussis and B. parapertussis; and (3) Verify the POC device for rapid detection of Bordetella species using clinical samples. The proposed methodology will allow for the confirmation of suspected cases of whooping cough caused by different Bordetella species at the point of care, filling a gap of current diagnostic methods in whooping cough. Furthermore, our biochip has great potential for the rapid detection of a variety of other respiratory infectious diseases such as COVID-19 in resource-poor settings.