A novel dynamic flow immunochromatographic test (DFICT) using gold nanoparticles for the serological detection of Toxoplasma gondii infection in dogs and cats

•A novel dynamic flow immunochromatographic test (DFICT) is developed.•A liquid gold nanoparticle conjugate reagent is introduced in DFICT.•It is not necessary to prepare the conjugated pad and the sample pad in DFICT.•DFICT requires only 5 μL of serum sample with no need for sample pretreatment.•DFICT can be used for serological detection of T. gondii in both dogs and cats.

A novel dynamic flow immunochromatographic test (DFICT) is proposed for rapid assay utilizing Toxoplasma gondii as a model. The test is based on a proprietary technology that combines the principles of immunochromatography and fluid dynamics. Gold nanoparticles conjugated to staphylococcal protein A (SPA) were prepared in liquid form and used as signal vehicles. T. gondii-specific recombinant antigens and SPA were sprayed onto a nitrocellulose membrane in strips at positions designated as T and C, respectively. The DFICT is performed by applying a 100 µL aliquot of liquid gold-SPA conjugate to the reagent hole and a 5 μL aliquot of serum sample to the sample hole. The results were observable within 5 min by the naked eye. The lowest detectable limit of the assay was determined as the highest dilution (1:320) of positive serum. No cross-reaction of the antibodies with other related canine or feline pathogens was observed. The DFICT can be stored for 12 months at 4 °C or 6 months with no loss of sensitivity or specificity. A high degree of consistency was observed between the DFICT and the standard ELISA kit, supporting the reliability of the novel test strip. The introduction of a liquid gold nanoparticle conjugate reagent provides this method with several attractive characteristics, such as ease of manufacture, low sample volume requirements, high selectivity and high efficiency. This method opens a novel pathway for rapid diagnostic screening and field analysis.