Enzyme-linked immunosorbent assay (ELISA) is a technique designed for the detection and quantification of (bio)­molecules in a liquid sample. It is a powerful tool in clinical diagnostics, food safety, and environmental monitoring. However, the main limitation of conventional ELISA is its low sensitivity, which cannot meet the demand of analyte analysis in complex (biological) samples. Several successful approaches capitalizing on the unique physical and chemical properties of nanoparticles to improve the performance of traditional ELISA have been reported. In this review, we aim to demonstrate diverse strategies designed to date that use metal and metal oxide nanoparticles to overcome challenges associated with ELISA sensitivity and stability. In particular, we discuss metal and metal oxide nanoparticles as carriers to load enzymes and antibodies for signal amplification, as enzyme mimics to replace the natural enzyme label, and as signal transducers to provide fluorescence or luminescence signals as an alternative output.

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Sensitivity is the key in optical detection of low-abundant analytes, such as circulating RNA or DNA. The enzyme Exonuclease III (Exo III) is a useful tool in this regard; its ability to recycle target DNA molecules results in markedly improved detection sensitivity. Lower limits of detection may be further achieved if the detection background of autofluorescence can be removed. Here we report an ultrasensitive and specific method to quantify trace amounts of DNA analytes in a wash-free suspension assay. In the presence of target DNA, the Exo III recycles the target DNA by selectively digesting the dye-tagged sequence-matched probe DNA strand only, so that the amount of free dye removed from the probe DNA is proportional to the number of target DNAs. Remaining intact probe DNAs are then bound onto upconversion nanoparticles (energy donor), which allows for upconversion luminescence resonance energy transfer (LRET) that can be used to quantify the difference between the free dye and tagged dye (energy acceptor). This scheme simply avoids both autofluorescence under infrared excitation and many tedious washing steps, as the free dye molecules are physically located away from the nanoparticle surface, and as such they remain "dark" in suspension. Compared to alternative approaches requiring enzyme-assisted amplification on the nanoparticle surface, introduction of probe DNAs onto nanoparticles only after DNA hybridization and signal amplification steps effectively avoids steric hindrance. Via this approach, we have achieved a detection limit of 15 pM in LRET assays of human immunodeficiency viral DNA.

The detection of pathogenic bacteria is essential to prevent and treat infections and to provide food security. Current gold-standard detection techniques, such as culture-based assays and polymerase chain reaction, are time-consuming and require centralized laboratories. Therefore, efforts have focused on developing point-of-care devices that are fast, cheap, portable and do not require specialized training. Paper-based analytical devices meet these criteria and are particularly suitable to deployment in low-resource settings. In this Review, we highlight paper-based analytical devices with substantial point-of-care applicability for bacteria detection and discuss challenges and opportunities for future development.

We report the synthesis of a catalyst, copper-doped zeolitic imidazolate framework ZIF-8, that generates nitric oxide from naturally occurring endogenous nitric oxide donors, S-nitrosoglutathione and S-nitrosocysteine.