We present the application of a photonic silicon chip-based optical sensor system for expeditious and phenotypic antifungal susceptibility testing. This label-free diagnostic assay optically monitors the growth of Candida auris at varying antifungal concentrations on a microwell-structured silicon chip in real-time, and antifungal susceptibility is detected within 6 h, four times faster than in the current gold standard method.
Analytical microbiology has made substantial progress since its conception, starting from potato slices, through selective agar media, to engineered surfaces modified with capture probes. While the latter represents the dominant approach in designing sensors for bacteria detection, the importance of sensor surface properties is frequently ignored. Herein, we highlight their significant role in the complex process of bacterial transition from planktonic to sessile, representing the first and critical step in bacteria detection. We present the main surface features and discuss their effect on the bio-solid interface and the resulting sensing capabilities for both flat and particulate systems. The concepts of rationally-designed surfaces for enhanced bacterial detection are presented with recent examples of sensors (capture probe-free) relying solely on surface cues.
The extensive and improper use of antibiotics has led to a dramatic increase in the frequency of antibiotic resistance among human pathogens, complicating infectious disease treatments. In this work, a method for rapid antimicrobial susceptibility testing (AST) is presented using microstructured silicon diffraction gratings integrated into prototype devices, which enhance bacteria-surface interactions and promote bacterial colonization. The silicon microstructures act also as optical sensors for monitoring bacterial growth upon exposure to antibiotics in a real-time and label-free manner via intensity-based phase-shift reflectometric interference spectroscopic measurements (iPRISM). Rapid AST using clinical isolates of Escherichia coli (E. coli) from urine is established and the assay is applied directly on unprocessed urine samples from urinary tract infection patients. When coupled with a machine learning algorithm trained on clinical samples, the iPRISM AST is able to predict the resistance or susceptibility of a new clinical sample with an Area Under the Receiver Operating Characteristic curve (AUC) of ∼ 0.85 in 1 h, and AUC > 0.9 in 90 min, when compared to state-of-the-art automated AST methods used in the clinic while being an order of magnitude faster.
The dominance of antibodies in diagnostics has gradually changed following the discovery of aptamers in the early 1990s. Aptamers offer inherent advantages over traditional antibodies, including higher specificity, higher affinity, smaller size, greater stability, ease of manufacture, and low immunogenicity, rendering them the best candidates for point-of-care testing (POCT). In the past 20 years, the research community and pharmaceutical companies have made great efforts to promote the development of aptamer technology. Macugen® (pegaptanib) was the first aptamer drug approved by the US Food and Drug Administration (FDA), and various aptamer-based diagnostics show great promise in preclinical research and clinical trials. In this review, we introduce recent literature, ongoing clinical trials, commercial reagents of aptamer-based diagnostics, discuss the FDA regulatory mechanisms, and highlight the prospects and challenges in translating these studies into viable clinical diagnostic tools.
Since oxalate plays an important role in the metabolic assessment of urolithiasis, there is need for convenient and efficient methods for oxalate detection. Herein, we report a three-signal fluorescence strategy for oxalate analysis based on the ability of oxalate to reduce Cu2+ to Cu+, and the ability of pyrophosphate-cerium coordination polymeric networks (PPi-Ce CPNs), CdTe quantum dots (QDs), and N-Methyl Mesoporphyrin (NMM) to selectively detect Cu2+ and Cu+. The detection range was 100 nM to 1 mM, the turnaround time was 6 min, while the limits of detections (LODs) for PPi-Ce CPNs, QDs and NMM as reporters were 25 nM, 10 nM and 40 nM, respectively. Visual detection of oxalate relied on color change in the solution, which could be observed using the naked eye. The fluorescent system was used for oxalate analysis in 44 urine samples (32 calcium oxalate stone patients, 12 controls without urolithiasis), and the results were consistent with clinical diagnosis and imaging data. Moreover, the visual system was used to analyze 8 urine samples (4 patients and 4 controls), and also showed good consistency with clinical diagnosis and computed tomography (CT) imaging results. These findings suggest that the method has potential application in the development of portable urolithiasis diagnosis kits.
With new advances in infectious disease, antifouling surfaces, and environmental microbiology research comes the need to understand and control the accumulation and attachment of bacterial cells on a surface. Thus, we employ intrinsic phase-shift reflectometric interference spectroscopic measurements of silicon diffraction gratings to non-destructively observe the interactions between bacterial cells and abiotic, microstructured surfaces in a label-free and real-time manner. We conclude that the combination of specific material characteristics (i.e., substrate surface charge and topology) and characteristics of the bacterial cells (i.e., motility, cell charge, biofilm formation, and physiology) drive bacteria to adhere to a particular surface, often leading to a biofilm formation. Such knowledge can be exploited to predict antibiotic efficacy and biofilm formation, and enhance surface-based biosensor development, as well as the design of anti-biofouling strategies.
Since the invention of the first biosensors 70 years ago, they have turned into valuable and versatile tools for various applications, ranging from disease diagnosis to environmental monitoring. Traditionally, antibodies have been employed as the capture probes in most biosensors, owing to their innate ability to bind their target with high affinity and specificity, and are still considered as the gold standard. Yet, the resulting immunosensors often suffer from considerable limitations, which are mainly ascribed to the antibody size, conjugation chemistry, stability, and costs. Over the past decade, aptamers have emerged as promising alternative capture probes presenting some advantages over existing constraints of immunosensors, as well as new biosensing concepts. Herein, we review the employment of antibodies and aptamers as capture probes in biosensing platforms, addressing the main aspects of biosensor design and mechanism. We also aim to compare both capture probe classes from theoretical and experimental perspectives. Yet, we highlight that such comparisons are not straightforward, and these two families of capture probes should not be necessarily perceived as competing but rather as complementary. We, thus, elaborate on their combined use in hybrid biosensing schemes benefiting from the advantages of each biorecognition element.