Abstract
In spite of a current increasing trend in the development of miniaturized, standalone point-of-care (PoC) biosensing platforms in the literature, the actual implementation of such systems in the field is far from being a reality although deeply needed. In the particular case of the population screenings for local or regional diseases related to specific pathogens, the diagnosis of the presence of specific antibodies could drastically modify therapies and even the organization of public policies. The aim of this work was to develop a fast, cost-effective detection method based on the manipulation of functionalized magnetic beads for an efficient diagnosis of hypersensitivity pneumonitis (HP), looking for the presence of anti-pigeon antigen antibodies (APAA) in a patient’s serum. We presented a Diagnostic Biosensor Method (DBM) in detail, with validation by comparison with a traditional high-throughput platform (ELISA assay). We also demonstrated that it was compatible with a microfluidic chip that could be eventually incorporated into a PoC for easy and broad deployment using portable optical detectors. After standardization of the different reaction steps, we constructed and validated a plastic chip that could easily be scaled to high-volume manufacturing in the future. The solution proved comparable to conventional ELISA assays traditionally performed by the clinicians in their laboratory and should be compatible with other antibody detection directly from patient samples.
Highlights
Hypersensitivity pneumonitis (HP) is a complex syndrome caused by exposure to a large variety of organic particles, small enough (~5 μm) to reach pulmonary alveoli [1,2]
We determined the concentration of pigeon sera (PS) for functionalization and the incubation time that allowed the capture of the greatest amount of IgG anti-pigeon antigen antibodies (APAA) in the samples
In the Diagnostic Biosensor Method (DBM) steps 2 and 3, serum antibodies from an hypersensitivity pneumonitis (HP)-positive patient (18.3 μg/80 K beads) and secondary antibody αHIgG-488 (1.7 μg/80 K beads) were used; both reactions had incubation times of 2 h and with 1.5 mL low-binding microtubes as the reaction platform
Summary
Hypersensitivity pneumonitis (HP) is a complex syndrome caused by exposure to a large variety of organic particles, small enough (~5 μm) to reach pulmonary alveoli [1,2]. HP is an immune-mediated interstitial lung disease occurring in susceptible individuals [1,2,3]. In a number of cases, HP can lead to pulmonary fibrosis, which is associated with progressive respiratory insufficiency and death. At work, at home, or even outdoors in recreational environments. In Mexico and many other countries, the main disease-associated exposure is contact with pigeons and domestic birds [4]. Key actions in HP treatment consist in early diagnosis and antigen avoidance, as it may allow limiting an aggressive progression of the disease in affected patients [5]
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