Rapid, label-free genetic detection of enteropathogens in stool without genetic isolation or amplification

Abstract

Current genetic detection methods require gene isolation, gene amplification and detection with a fluorescent-tagged probe. They typically require sophisticated equipment and expensive fluorescent probes, rendering them not widely available for rapid acute infection diagnoses at the point of care to ensure timely treatment of the diseases. Here we report a rapid genetic detection method that can detect the bacterial gene directly from patient stools using a piezoelectric plate sensor (PEPS) in conjunction with a continuous flow system with two temperature zones. With stools spiked with sodium dodecyl sulfate (SDS) in situ bacteria lysing and DNA denaturation occurred in the high-temperature zone whereas in situ specific detection of the denatured DNA by the PEPS occurred in the lower-temperature zone. The outcome was a rapid genetic detection method that directly detected bacterial genes from stool in <40 min without the need of gene *

Sensitive Multi‐Detection of Escherichia coli by Quartz Crystal Microbalance with a Novel Surface Controllable Sensing Method in Liquid Organic Fertilizer Produced by Sewage Sludge

“…When FMSs functionalized with antibodies that specifically capture surface antigens of E. coli bacteria are applied to the sensor surface, the modified FMSs bind to the target bacteria that have been captured on the sensor surface by the same antibody‐antigen interaction. The mass increase, which cannot be obtained with a small number of binding of the bacteria in the low‐concentration sample to the surface, is achieved by providing an extra mass increase with FMSs [42] that can bind specifically to the target.…”

Visual and Rapid Detection of Escherichia coli O157:H7 in Stool Samples by FTA Card-based Loop-mediated Isothermal Amplification

“…Compared to the gold standard method (PCR) for the detection of E. coli O157:H7 from stool samples, the assay we developed not only achieved higher sensitivity (25 CFU/mL) in a shorter time (35 min) without complex sample preparation, operation, reaction, and analysis steps, but was also tolerant to PCR inhibitors and reduced interference to the reaction [18,21]. Remarkably, our approach only requires a compact, portable, and lightweight dry bath, eliminating the requirement for cumbersome and complicated equipment.…”

Label-Free Biosensors for Laboratory-Based Diagnostics of Infections: Current Achievements and New Trends

“…Moreover, an important advantage of the use of label-free biosensors is that target analytes are detected in their natural form without labeling and chemical modification, and thus, they can be preserved for further analysis (Table 2). [ 22,26,39,80,86] In recent decades, numerous studies have been carried out to develop new types of receptors [29,63,87,88] and methods of recognition for label-free biosensors. They can generate signal immediately after binding to the recognition element and do not require additional interactions with the labels that provide signal.…”

“…Due to the ease of use and high sensitivity, electrochemical and optical label-free biosensors have been most commonly deployed for specific and non-specific indication of biological agents in recent years (Table 3). Monoclonal antibodies Visualization Salmonella enteritidis, 10 2 -10 8 ufc/mL [88] DNA, aptamer Electrochemical Bird flu virus H5N1 (AIV) [48] Nucleic acids (DNA) Electrochemical impedance Zika virus, 25.0 ± 1.7 hM.…”