Rapid and Ultrasensitive Colorimetric Biosensors for Onsite Detection of Escherichia coli O157:H7 in Fluids.

This study presents a breakthrough in the field of onsite bacterial detection, offering an innovative, rapid, and ultrasensitive colorimetric biosensor for the detection of Escherichia coli (E. coli) O157:H7, using chemically modified melamine foam (MF). Different from conventional platforms, such as 96-well plates and fiber-based membranes, the modified MF features a macroporous reticulated three-dimensional (3D) framework structure, allowing fast and free movement of large biomolecules and bacteria cells through the MF structure in every direction and ensuring good accessibility of entire active binding sites of the framework structure with the target bacteria, which significantly increased sensitive and volume-responsive detection of whole-cell bacteria. The biosensing platform requires less than 1.5 h to complete the quantitative detection with a sensitivity of 10 cfu/mL, discernible by the naked eye, and an enhanced sensitivity of 5 cfu/mL with the help of a smartphone. Following a short enrichment period of 1 h, the sensitivity was further amplified to 2 cfu/mL. The biosensor material is volume responsive, making the biosensing platform sensitivity increase as the volume of the sample increases, and is highly suitable for testing large-volume fluid samples. This novel material paves the way for the development of volume-flexible biosensing platforms for the record-fast, onsite, selective, and ultrasensitive detection of various pathogenic bacteria in real-world applications.

Discovery of Peptidic Ligands against the SARS-CoV-2 Spike Protein and Their Use in the Development of a Highly Sensitive Personal Use Colorimetric COVID-19 Biosensor.

In addition to efficacious vaccines and antiviral therapeutics, reliable and flexible in-home personal use diagnostics for the detection of viral antigens are needed for effective control of the COVID-19 pandemic. Despite the approval of several PCR-based and affinity-based in-home COVID-19 testing kits, many of them suffer from problems such as a high false-negative rate, long waiting time, and short storage period. Using the enabling one-bead-one-compound (OBOC) combinatorial technology, several peptidic ligands with a nanomolar binding affinity toward the SARS-CoV-2 spike protein (S-protein) were successfully discovered. Taking advantage of the high surface area of porous nanofibers, immobilization of these ligands on nanofibrous membranes allows the development of personal use sensors that can achieve low nanomolar sensitivity in the detection of the S-protein in saliva. This simple biosensor employing naked-eye reading exhibits detection sensitivity comparable to some of the current FDA-approved home detection kits. Furthermore, the ligand used in the biosensor was found to detect the S-protein derived from both the original strain and the Delta variant. The workflow reported here may enable us to rapidly respond to the development of home-based biosensors against future viral outbreaks.

Improving the Sensitivity of Nanofibrous Membrane-Based ELISA for On-Site Antibiotics Detection.

An ultrasensitive and portable colorimetric enzyme-linked immunosorbent assay (ELISA) sensor for antibiotics was fabricated by immobilizing antibodies inside the largely porous and highly hydrophilic nanofibrous membranes. Different from regular electrospun nanofibrous membranes where antibodies may frequently be blocked by the heterogeneous porous structure and sterically crowded loaded on the surface, the controlled microporous structure and increased hydrophilicity of nanofibrous membranes could improve the diffusion properties of antibodies, reduce the sterically crowding effect, and dramatically improve the sensitivity of the membrane-based ELISA. The limitation of detection (LOD) for chloramphenicol (CAP) reached 0.005 ng/mL, around 200 times lower than the conventional paper-based ELISA, making quantitative analysis and portable on-site detection achievable via the use of smartphones. The successful design and fabrication of the nanofibrous membrane-based ELISA with novel features overcome the structural drawbacks of regular electrospun nanofibrous membranes and provide new paths to develop highly sensitive on-site detection of hazardous chemical agents.

Photoactivities of Two Vitamin B Derivatives and Their Applications in the Perpetration of Photoinduced Antibacterial Nanofibrous Membranes.

Photoactivities and photoinduced antibacterial functions of two vitamin B2 (VB2) derivatives, riboflavin (RF) and flavin mononucleotide (FMN), were investigated by computational modeling and various experimental evaluations. Under photoirradiation, the ground state of both VB2 derivatives could be excited to generate different reactive oxygen species (ROS) by undergoing different reaction paths. The formed ROS could nonselectively inactivate microorganisms. However, both RF and FMN exhibited negligible photoinduced antimicrobial activity in phosphate-buffered saline (PBS) solutions even at high concentrations. The study revealed that the affinity of both RF and FMN to microorganisms in different application media plays a key role due to the very short lifetime of the generated ROS. The speculation was proven by the preparation of a poly(vinyl alcohol-co-ethylene) (PVA-co-PE) nanofibrous membrane blended with RF or FMN, which could enhance the contact of the agents with microorganisms. The fabricated nanofibrous membranes containing both VB2 derivatives (VBNFMs) showed great photoinduced antibacterial activity against Gram-negative Escherichia coli (E. coli) (99.999% bacterial reduction) and Gram-positive Listeria innocua (L. innocua) (99% bacterial reduction) under 20 min of ultra-violet A irradiation. The photoinduced antimicrobial performances of RF/PVA-co-PE and FMN/PVA-co-PE nanofibrous membranes were comparable. Interestingly, the durability of the photoinduced antibacterial functions of the prepared VBNFMs was questionable, due to the photodegradation of VB2 in nanomaterials.

Chlorine Rechargeable Biocidal N-Halamine Nanofibrous Membranes Incorporated with Bifunctional Zwitterionic Polymers for Efficient Water Disinfection Applications.

An intrinsically hydrophilic nanofibrous membrane with chlorine rechargeable biocidal and antifouling functions was prepared by using a combination of chemically bonded N-halamine moieties and zwitterionic polymers (PEI-S). The designed nanofibrous membrane, named as PEI-S@BNF-2 h, can exhibit integrated features of reduced bacterial adhesion, rechargeable biocidal activity, and easy release of killed bacteria by using mild hydrodynamic forces. The representative functional performances of the PEI-S@BNF-2 h membrane include high active chlorine capacity (>4000 ppm), large specific surface area, ease of chlorine rechargeability, long-term stability, and exceptional biocidal activity (99.9999% via contact killing). More importantly, the zwitterionic polymer moieties (PEI-S) brought robust antifouling properties to this biocidal membrane, therefore reducing the biofouling-biofilm effect and prolonging the lifetime of the filtration membrane. These attributes enable the PEI-S@BNF-2 h nanofibrous membrane to effectively disinfect the microbe-contaminated water with high fluxes (10,000 L m-2 h-1) and maintain itself clean for a long-term application.

Design and fabrication of a highly sensitive and naked-eye distinguishable colorimetric biosensor for chloramphenicol detection by using ELISA on nanofibrous membranes.

Enzyme-linked immunoassay (ELISA) is highly specific and selective towards target molecules and is convenient for on-site detection. However, in many cases, lack of high sensitivity makes it hard to reveal a significant colorimetric signal for detecting a trace amount of target molecules. Thus, analytical instruments are required for detection, which limits the application of ELISA for on-site detection. In the present study, a highly sensitive and naked-eyed detectable colorimetric biosensor for chloramphenicol (CAP) was prepared by incorporating ELISA onto surfaces of microporous and nanofibrous membranes. The high specific surface areas of the nanofibers significantly increased the number of antibodies covalently linked onto the fiber surfaces and binding capacity of the sensor with antigens present in a sample. With such an integration, the sensitivity of the ELISA sensor was dramatically increased, and a trace number of targets could reveal a naked-eye detectable color. The immunoassay sensor exhibited a significant naked-eye distinguishable color to chloramphenicol (CAP) at 0.3 ng/mL. The successful design and fabrication of the nanofibrous membrane immunoassay sensor provide new paths towards the development of on-site inspection sensors without the assistance from any instrument.