Paper-Based Test for Rapid On-Site Screening of SARS-CoV-2 in Clinical Samples

Detection methods for monitoring infectious pathogens has never been more important given the need to contain the spread of the COVID-19 pandemic. Herein we propose a highly sensitive magnetic-focus-enhanced lateral flow assay (mLFA) for the detection of SARS-CoV-2. The proposed mLFA is simple and requires only lateral flow strips and a reusable magnet to detect very low concentrations of the virus particles. The magnetic focus enhancement is achieved by focusing the SARS-CoV-2 conjugated magnetic probes in the sample placed in the lateral flow (LF) strips for improved capture efficiency, while horseradish peroxidase (HRP) was used to catalyze the colorimetric reaction for the amplification of the colorimetric signal. With the magnetic focus enhancement and HRP-based amplification, the mLFA could yield a highly sensitive technology for the recognition of SARS-CoV-2. The developed methods could detect as low as 400 PFU/mL of SARS-CoV-2 in PBS buffer based on the visible blue dots on the LF strips. The mLFA could recognize 1200 PFU/mL of SARS-CoV-2 in saliva samples. With clinical nasal swab samples, the proposed mLFA could achieve 66.7% sensitivity and 100% specificity.

On-Site Detection of Carcinoembryonic Antigen in Human Serum

Real-time connectivity and employment of sustainable materials empowers point-of-care diagnostics with the capability to send clinically relevant data to health care providers even in low-resource settings. In this study, we developed an advantageous kit for the on-site detection of carcinoembryonic antigen (CEA) in human serum. CEA sensing was performed using cellulose-based lateral flow strips, and colorimetric signals were read, processed, and measured using a smartphone-based system. The corresponding immunoreaction was reported by polydopamine-modified gold nanoparticles in order to boost the signal intensity and improve the surface blocking and signal-to-noise relationship, thereby enhancing detection sensitivity when compared with bare gold nanoparticles (up to 20-fold in terms of visual limit of detection). Such lateral flow strips showed a linear range from 0.05 to 50 ng/mL, with a visual limit of detection of 0.05 ng/mL and an assay time of 15 min. Twenty-six clinical samples were also tested using the proposed kit and compared with the gold standard of immunoassays (enzyme linked immunosorbent assay), demonstrating an excellent correlation (R = 0.99). This approach can potentially be utilized for the monitoring of cancer treatment, particularly at locations far from centralized laboratory facilities.

Rapid detection of mcyG gene of microcystins producing cyanobacteria in water samples by recombinase polymerase amplification combined with lateral flow strips

Nowadays, cyanobacteria blooms and microcystins (MCs) pollution are threatening water safety and public health. In this study, a rapid detection method was established for detecting MCs producing cyanobacteria. The MC synthesis gene mcyG was measured through recombinase polymerase amplification combined with lateral flow strips (LF-RPA) technology. The target gene mcyG was amplified at a temperature range of 37–45 °C, and the amplification time to detect mcyG was only 15 min at 37 °C. The optimal reaction conditions were confirmed using single dependent variable experiments, suggesting that the best probe dosage for 50 μL of the reaction mixture was 0.2 μL, the best dilution ratio of products was 1/100, and the best loading volume was 10 μL. The specificity test proved that the LF-RPA assay could distinguish MCs producing cyanobacteria from nontoxic algae well. Within 35 min of amplification time, the detection limit of the LF-RPA assay was 103 copies/mL mcyG and 104 cells/mL Microcystis aeruginosa FACHB-905. Overall, the LF-RPA assay could detect MCs producing cyanobacteria in water samples quickly and accurately, and it has a great promise to be applied for monitoring the MCs producing cyanobacteria blooms in natural waters.