Current and emerging molecular diagnostic technologies applicable to bacterial food safety

There is an increasing need for rapid test methods to certify the quality and safety of food products. Current tests applied for the microbiological assessment of food products are based on standard approved culture-based isolation methods and can take several days to yield results. Nucleic acid diagnostic (NAD) tests for the identification of bacterial foodborne pathogens employing in vitro amplification technologies are capable of sensitive and specific detection of single or multiple pathogens in foods in a shorter timeframe than traditional methods. New developments in molecular biosensors have the potential to provide at-line bioanalysis, whereas microarray-based technologies may in the future be the NAD platforms of choice for multiple pathogen detection and identification. This article reviews current and emerging NAD platforms for foodborne bacterial pathogens that have the potential to impact food safety.     

Synthesis,Functionalization, and Bioconjugation of Monodisperse,Silica‐Coated Gold Nanoparticles: Robust Bioprobes

Herein, we report an efficient process for preparing monodisperse Au@SiO₂ nanoparticles using homogeneous shaking and without the use of surface‐coupling silane agents or large stabilizers. The resulting pure‐silica surface of the Au@SiO₂ nanoparticles is very important for straightforward surface functionalization with different functional groups via well‐established silica surface chemistry. Subsequent covalent bioconjugation of the aldehyde‐functionalized Au@SiO₂ nanoparticles with various biomolecules is successfully employed to make robust nanoprobes for fast, colorimetric DNA and protein detection based on the sequence‐specific hybridization properties of DNA and the specific binding affinity between proteins, respectively.     

Study on design and application of fully automatic miniature surface plasmon resonance concentration analyzer

A fully automatic miniature surface plasmon resonance (SPR) concentration analyzer having high performance and low cost and developed using a Spreeta™ sensor was designed for field applications and concentration analysis. As in the case of Biacore™ instruments, the automatic sampling system of this device can introduce air segments between the sample/regeneration solution and buffer solution in the pipeline, which effectively prevents mixing of the solutions. A temperature sensor (AD 590) and temperature compensation method are used, which make the device insensitive to temperature fluctuations. A real-time data-smoothing algorithm for the SPR detection data is adopted; this can reduce the noise level to 5 × 10⁻⁷ RIU (refractive index units). The noise level of the sensorgram is 3.5% of the original level. Two types of self-prepared sensing chips—SMX-BSA (bovine serum albumin coated with sulfamethoxazole) and SMX-CM5 (carboxymethyl dextran coated with sulfamethoxazole)—are used to analyze the concentrations of sulfamethoxazole (SMX) standard solutions. Each chip's SMX calibration curve is established within the measurement range of 0-2000 ng/ml, and both limits of detection (LOD) are 2 ng/ml. One cycle of assay time is less than 15 min.     

Micro-piezoelectric immunoassay chip for simultaneous detection of Hepatitis B virus and α-fetoprotein

In this paper, a piezoelectric diaphragm-based immunoassay chip was developed to simultaneously detect anti-Hepatitis B virus (HBV) and anti-alpha-fetoprotein (AFP). The chip was fabricated by micro-machining technology and consists of eight individual circular sensors with a diameter of 800 μm. Hepatitis B surface antigen (HBsAg), Hepatitis C core antigen (HBcAg) and AFP as the probe molecules were immobilized on different sensing spots on the chip. A solution containing anti-HBsAg and anti-AFP was applied into the reaction chambers in all sensors of the chip, and significant frequency shifts were only observed in the sensors with HBsAg and AFP for immunoassay detection. The fluorescence image further confirmed the successful detection of anti-HBsAg and anti-AFP. The total assay time was less than 2 h. The frequency shift-based calibration curves show a detection limit of 0.1 ng/ml and a dynamic detection range of 0.1-10,000 ng/ml for both anti-HBsAg and anti-AFP, respectively, thus demonstrating that the developed piezoelectric immunoassay chip has potential applications for rapid, specific, sensitive, and multiple detections of HBV.     

A MEMS affinity glucose sensor using a biocompatible glucose-responsive polymer

We present a MEMS affinity sensor that can potentially allow long-term continuous monitoring of glucose in subcutaneous tissue for diabetes management. The sensing principle is based on detection of viscosity changes due to affinity binding between glucose and poly(acrylamide-ran-3-acrylamidophenylboronic acid) (PAA-ran-PAAPBA), a biocompatible, glucose-specific polymer. The device uses a magnetically driven vibrating microcantilever as a sensing element, which is fabricated from Parylene and situated in a microchamber. A solution of PAA-ran-PAAPBA fills the microchamber, which is separated from the surroundings by a semi-permeable membrane. Glucose permeates through the membrane and binds reversibly to the phenylboronic acid moiety of the polymer. This results in a viscosity change of the sensing solution, which is obtained by measuring the damped cantilever vibration using an optical lever setup, allowing determination of the glucose concentration. Experimental results demonstrate that the device is capable of detecting glucose at physiologically relevant concentrations from 27 mg/dL to 324 mg/dL. The glucose response time constant of the sensor is approximately 3 min, which can be further improved with device design optimization. Excellent reversibility and stability are observed in sensor responses, as highly desired for long-term, stable continuous glucose monitoring.     

Cover Picture: Protein Detecting Arrays Based on Cationic Polythiophene–DNA‐Aptamer Complexes (Adv. Mater. 20/2006)

The cover image depicts biochips based on responsive nanoaggregates made from stoichiometric complexes between a cationic polythiophene and an appropriate DNA aptamer. These structures undergo a conformational transition from an unfolded to a folded (G‐quadruplex) structure in the presence of a specific target protein that results in a significant increase of the fluorescence intensity, as reported on p. 2703 by Leclerc and co‐workers.