Background: Gold nanoparticle-based immunochromatographic assay (AuNP-ICA) has insufficient sensitivity due to its inherent colorimetric signal intensity and low capture efficiency of AuNPs. The metal in situ growth is a common strategy to enhance the sensitivity of AuNP-ICA due to its superior signal amplification potential and simple operation. However, the detection distortion caused by metal self-nucleation during the growth process can seriously affect the accuracy and reproducibility of the strips. Methods: We present a pH-regulated gold in situ growth (GISG) strategy to amplify the colorimetric signal and demonstrate its application in improving the performance of traditional AuNP-ICA. The controllable growth signal amplification is achieved by lowering the pH of the growth solution to weaken the reducibility of hydroxylamine (HA), thus urging the crystallization and growth of Au 3+ on the AuNP surface instead of free reduction and self-nucleation. In addition, the mechanism of pH regulation on HA reducibility is elucidated by introducing an electron-donating or electron-withdrawing group to affect the electron density of hydroxyl group. Results: The proposed GISG strategy shows improved sensitivity, low background, robust operation, and good reproducibility. The LOD values of the designed GISG-amplified AuNP-ICA are as low as 0.0198 ng mL -1 for hepatitis B surface antigen and 0.0125 ng mL -1 for HIV-1 capsid p24 antigen, which are lower by about 500- and 70-fold, respectively, than those of the unamplified AuNP-ICA. Conclusions: This method is extended to enable ultrasensitive and rapid diagnosis of viral infections, and has potential as a general signal amplification platform to redefine immunochromatographic diagnostics.

We applied urease-induced silver metallization on the surface of gold nanorods (AuNR) to improve colorimetric ELISA for the rapid and sensitive detection of Salmonella enterica Choleraesuis. To this end, we introduced a biotin-streptavidin system as a bridge to determine the correlation between urease and S. enterica Choleraesuis concentrations. The captured urease can catalyze the hydrolysis of urea into carbon dioxide and ammonia, and the generated ammonia can then induce the deposition of silver shell on the surface of AuNR in the presence of silver nitrate and glucose. With the decreased aspect ratio (length divided by width) of AuNR, a multicolor change of AuNR solution and a significant blue shift in the longitudinal localized surface plasmon resonance absorption peak (∆λ max) of AuNR were obtained at the target concentrations of 1.21 × 10 1 to 1.21 × 10 8 cfu/mL. Consequently, the detection limits of our proposed colorimetric ELISA were as low as 1.21 × 10 2 cfu/mL for qualitative detection with naked eyes, and 1.21 × 10 1 cfu/mL for quantitative detection, in which changes in ∆λ max of AuNR were recorded with a microplate reader. These values were at least 2 to 3 orders of magnitude lower than those obtained with conventional horseradish peroxidase-based ELISA. The analytical performance of our developed colorimetric ELISA in terms of selectivity, accuracy, reliability, and practicability were investigated by analyzing S. enterica Choleraesuis-spiked pasteurized whole milk samples.

We report the preparation and application of biofunctional nanoparticles to detect C-reactive protein (CRP) in magnetic microplates. A CRP model biomarker was used to test the proposed detection method. Biofunctional magnetic nanoparticles, CRP, and biofunctional fluorescent nanoparticles were used in a sandwich nanoparticle immunoassay. The CRP concentrations in the samples were deduced from the reference plot, using the fluorescence intensity of the sandwich nanoparticle immunoassay. When biofunctional nanoparticles were used to detect CRP, the detection limit was 1.0 ng ml(-1) and the linear range was between 1.18 ng ml(-1) and 11.8 μg ml(-1). The results revealed that the method involving biofunctional nanoparticles exhibited a lower detection limit and a wider linear range than those of the enzyme-linked immunosorbent assay (ELISA) and most other methods. For CRP measurements of serum samples, the differences between this method and ELISA in CRP measurements of serum samples were less than 13%. The proposed method can reduce the analysis time to one-third that of ELISA. This method demonstrates the potential to replace ELISA for rapidly detecting biomarkers with a low detection limit and a wide dynamic range.