Detection of isothermally amplified ostreid herpesvirus 1 DNA in Pacific oyster (Crassostrea gigas) using a miniaturised electrochemical biosensor
TALANTA
Authors: Toldra, Anna; Dolors Furones, M.; O'Sullivan, Ciara K.; Campas, Monica
Abstract
Given the threat that ostreid herpesvirus 1 (OsHV-1) poses to shellfish aquaculture, the need for rapid, user-friendly and cost-effective methods to detect this marine pathogen and minimise its impact is evident. In this work, an electrochemical biosensor for the detection of OsHV-1 based on isothermal recombinase polymerase amplification (RPA) was developed. The system was first tested and optimised on maleimide microtitre plates as a proof-of-concept, before being implemented on miniaturised gold electrodes. Amperometric detection of the isothermally amplified product was achieved through a sandwich hybridisation assay with an immobilised thiolated capture probe and a horseradish peroxidase (HRP)-labelled reporter probe. Calibration curves were constructed using PCR-amplified OsHV-1 DNA, achieving a limit of detection of 207 OsHV-1 target copies. The biosensor was applied to the analysis of 16 oyster samples from an infectivity experiment, and results were compared with those obtained by qPCR analysis, showing a strong degree of correlation (r = 0.988). The simplicity, rapidity, cost-effectiveness and potential for in-situ testing with the developed biosensor provide a valuable tool for the detection of OsHV-1 in aquaculture facilities, improving their management.
Application of Horseradish Peroxidase Labeling Along With Post-Staining Improves Data Quality in Micro-Brain Connectomics
PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
Authors: Wang Sheng-Xiong; Chen Zhen-Qiang; Han Hua; Guo Ai-Ke
Abstract
For micro-brain connectomic research, volume electron microscopy is a main method in which it plays an importatnt role for the appliaction of automated tape-collecting ultramicrotome (ATUM) system and arrayimaging with scanning electron microscopy together. ATUM system enables collecting a large amount of special sections to be imaged with SEM to form ahuge dataset. With regard tothe nature of non-vacuum sectoningand sample reserving. we modified and improved the sample preparatio process includingresinrecipe, HRP labelling and post-staing method i orderto reduce folding rate of sample sections and make it possible for identifyig certain types of neurons in high-contrast imaging dataset, thus obtainig dataset of high quality for connectomic research.