Rapid vertical flow technique for the highly sensitive detection of Brucella antibodies with Prussian blue nanoparticle labeling and nanozyme-catalyzed signal amplification.

Brucellosis is a chronic infectious disease caused by Brucella, which is characterized by inflammation of reproductive organs and fetal membranes, abortion, infertility, and local inflammatory lesions of various tissues. Due to the widespread prevalence and spread of brucellosis, it has not only caused huge losses to animal husbandry, but also brought serious impacts on human health and safety. Therefore, rapid and accurate diagnosis is of great significance for the effective control of brucellosis. Therefore, we have developed a rapid vertical flow technique (RVFT) using Prussian blue nanoparticles (PBNPs) as a marker material for the detection of brucellosis antibodies. Lipopolysaccharide (LPS) was purified and used to detect brucellosis antibodies to improve the sensitivity of this technique. To enhance the sensitivity of serum antibody detection, a single multifunctional compound buffer was created using whole blood as a biological sample while retaining the advantages of typical lateral flow immunoassays. After signal amplification, standard Brucella-positive serum (containing Brucella antibody at 4000 IU mL) could be detected in this system even at a dilution factor of 1 × 10. The detection limit was 40 IU mL, which is ten times that before signal amplification. This RVFT displayed good specificity and no cross-reactivity. This RVFT effectively avoided the false negative phenomenon of lateral flow immunoassays, was easy to operate, had a short reaction time, has good repeatability, and could elicit results that were visible to the naked eye for 2 ~ 3 min without any equipment. Since this method is very important for controlling the prevalence of brucellosis, it holds great promise for application in primary medical units and veterinary brucellosis detection.