A new immunosensor for the determination of procalcitonin was developed. A sandwich assay format was implemented on a polymethylmetacrylate optical biochip, opportunely shaped in order to obtain several flow channels and potentially suitable for point of care testing applications. The sandwich format makes use of two new rat monoclonal antibodies. The capture antibody was covalently immobilised on the surface of the plastic chip, and the detection antibody was labelled with DY647 dye. Different combinations of capture and detection antibodies were investigated, and particular attention was devoted in order to avoid the non-specific adsorption. A limit of detection of 0.088 mg L(-1) was achieved within the working range of 0.28-50 mg L(-1) in buffer samples. The assay was also implemented in human serum, and 0.2 and 0.7-25 mg L(-1) were the attained limit of detection and working range, respectively.

No abstract

The discrimination of viral and bacterial sepsis is an important issue in intensive care patients. For this purpose, the simultaneous measurements of different analytes are necessary. Among the possible candidates, C-reactive protein (CRP) and procalcitonin (PCT) are probably the most important ones. A novel optical platform was designed and realised for the implementation of fluorescence-based immunoassays. The core of the optical platform is a plastic biochip, constituted by 13 microchannels (50 μm high, 600 μm width, 10 mm long) through which the sample flows. The sensing layer, where the immunochemical reaction takes place, is located on the upper part of each microchannel. The chip is interrogated with a novel optoelectronic platform, based on fluorescence anisotropy. A line-shaped beam from a 635-nm laser-diode excites perpendicularly the sensing layer and great many of the emitted remains entrapped inside the chip. The particular shape of the top of the chip allows to guide the emitted fluorescence along the same direction of the microchannel. The fluorescence which comes out on the lateral side from the chip is collected by a single plastic optical fibre and sent to an amplified photodiode. The device was characterised by the implementation of the sandwich assay for CRP and PCT spiked in serum. Limit of quantifications of 4.5 and of 6 µg L -1 in serum solution were achieved for CRP and PCT, respectively.

No abstract

The discrimination of viral and bacterial sepsis is an important issue in intensive care patients. For this purpose, the simultaneous measurements of different analytes such as C-reactive protein (CRP), procalcitonin (PCT), myeloperoxidase, interleukines and neopterin, are necessary. A novel optical platform was designed and realised for the implementation of fluorescence-based immunoassays. The core of the optical platform is a plastic biochip, formed by a series of microchannels each of them devoted to the determination of a single analyte. Sandwich assays for CRP and PCT spiked in serum were performed in order to demonstrate the reliability of a multi-array device.