Fast and sensitive graphene oxide‐DNAzyme‐based biosensor for Vibrio alginolyticus detection

Abstract

DNAzymes have been widely and effectively used for the detection of pathogenic bacteria, which pose a serious public health threat. However, the rapid and cost-effective detection of such bacteria remains a major challenge. In this study, we successfully selected Vibrio alginolyticus-specific DNAzymes. The activity of the candidates was assessed via fluorescence intensity and gel electrophoresis. The DNAzyme DT1 had a detection limit of 31 CFU/ml for V. alginolyticus and exhibited high specificity.Graphene oxide (GO) was used to develop a DNAzyme-based fluorescent sensor for the detection of V. alginolyticus, which significantly improved detection performance and shortened the reaction time as little as 10 s. The proposed method was then validated using crab, shrimp, fish, clam, and oyster samples. This study thus provides a new method for the rapid and sensitive detection of V. alginolyticus.

Emerging technologies revolutionising disease diagnosis and monitoring in aquatic animal health

“…Gold nanoparticle biosensors have detected fish necrosis virus RNA with a sensitivity of 270 picograms (pg), 64 and graphene oxide-based sensors have been used to detect Vibrio alginolytics up to 31 colonyforming units (CFU/ml). 117 DNAzyme (D-AS-2) combined with graphene can detect bacterial disease caused by Aeromonas salmonicida, producing fluorescence within 2 min. 116 Some viral diseases, such as viral hemorrhagic septicemia (VHSV), cause significant losses in fish farming.…”

“…115 In aquaculture, biosensors are used to detect cyanobacteria, cyanobacterial toxins in water and disease diagnostic. 116,117 Biosensors have various applications in detecting stress and pathogens in fish. For example, a biosensor system was developed to detect cortisol in fish plasma samples as a stress indicator in Oreochromis niloticus.…”

Screening of Sialyllactose-Specific Aptamer and Engineering a Pair of Recognition Elements with Unique Fluorescent Characteristics for Sensitive Detection of Sialyllactose

“…The affinity of the candidate aptamers was characterized by dissociation constant (K d ) and determined by graphene oxide (GO)-based displacement method 1 . Briefly, 100 μL 5'-FAM-labeled aptamer sequences at different concentrations (200, 400, 600, 800, and 1000 nM) were mixed with 100 μL of 50 mM Tris-HCl binding buffer (pH 7.6), denatured at 90 ℃ for 10 min and cooled down to 0 ℃ for 10 min.…”

A Novel Method for Sensitive Detection of Vibrio alginolyticus Based on Aptamer and Hybridization Chain Reaction in Aquaculture

“…However, the long process and cumbersome operation cannot meet the requirements of rapid detection [10]. Several molecular biological techniques are also used to detect V. alginolyticus, such as polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA) based on molecular amplification technology, enzyme-linked immunosorbent assay (ELISA), and immunofluorescence assay (IFA) based on immunology [11][12][13]. The molecular amplification-based detection method, which involves DNA extraction and amplification, enables highly sensitive detection down to the single-cell level.…”