Frequent outbreaks of foodborne illness have been increasing the awareness of food safety. Conventional methods for pathogen detection and identification are labor-intensive and take days to complete. Some immunological, rapid assays are developed, but these assays still require prolonged enrichment...
Frequent outbreaks of foodborne illness have been increasing the awareness of food safety. Conventional methods for pathogen detection and identification are labor-intensive and take days to complete. Some immunological, rapid assays are developed, but these assays still require prolonged enrichment steps. Recently developed biosensors have shown potential for the rapid detection of foodborne pathogens. In this study, an impedimetric biosensor was developed for rapid detection of Salmonella entritidis in food sample. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using a semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on neutravidin-biotin binding on the surface of the IME to form an active sensing layer. To evaluate the effect of electrode gap on sensitivity of the sensor, 3 types of sensors with different electrode gap sizes (2, 5, and $10{\mu}m$) were fabricated and tested. The impedimetric biosensor could detect $10^3\;CFU/mL$ of Salmonella in pork meat extract with an incubation time of 5 min. This method may provide a simple, rapid, and sensitive method to detect foodborne pathogens.
Frequent outbreaks of foodborne illness have been increasing the awareness of food safety. Conventional methods for pathogen detection and identification are labor-intensive and take days to complete. Some immunological, rapid assays are developed, but these assays still require prolonged enrichment steps. Recently developed biosensors have shown potential for the rapid detection of foodborne pathogens. In this study, an impedimetric biosensor was developed for rapid detection of Salmonella entritidis in food sample. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using a semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on neutravidin-biotin binding on the surface of the IME to form an active sensing layer. To evaluate the effect of electrode gap on sensitivity of the sensor, 3 types of sensors with different electrode gap sizes (2, 5, and $10{\mu}m$) were fabricated and tested. The impedimetric biosensor could detect $10^3\;CFU/mL$ of Salmonella in pork meat extract with an incubation time of 5 min. This method may provide a simple, rapid, and sensitive method to detect foodborne pathogens.
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제안 방법
In this research, impedimetric biosensors for detecting S. enteritidis in food samples were developed using an interdigitated microelectrode-based immunosensor. The impedance magnitude of the biosensor was measured at frequency ranges between 100 Hz to 1 MHz with a 50 mV (amplitude) voltage excitation.
In this study, impedimetric biosensors were developed and evaluated for detection of S. enteritidis in food samples. To increase sensitivity of the biosensor, different specifications of interdigitated electi'odes were fabricated and tested.
Specification of the biosensor and binding of S. enteritidis to antibodies immobilized on the biosensor surface was examined with a scanning electron microscopy (SEM). Figure 3 shows the SEM image (l, 000x) of the biosensor before (A) and after (B) the detection assay.
Active sensing area was created by soaking the wafer in the acetone to lift off the metal not adhering to the grass substrate. Three different sensor types with different electrode gap sizes (2, 5, and 10 |im) were fabricated to evaluate the effect of electrode specification on the sensitivity of the sensor. Each sensor had a 3 mm2 active sensing area.
대상 데이터
For sample preparation, selenite broth was purchased from Sigma- Al&ich. Salmonella enterica serotype enteritidis was obtained from Dr. Bhunia's laboratory (Purdue University, IN, USA) and used for the experiments. The bacteria were maintained on brain heart infusion (BHI) agar (1.
Each sensor had a different number of electrodes according to electrode spacing. Total numbers of electrodes were 250, 200, and 150 for 2, 5, and 10 μm of gap size, respectively. The IME sensor design is shown in Fig.
이론/모형
For negative control, PBS or plain food extract, which did not contain Salmonella cells, was used. Enumeration of the enriched S. enteritidis was performed using the standard plate count (SPC) method. For the food sample preparation, packages of pork were purchased from a local grocery store.
The IME sensor was fabricated by using a photolithographic processing method. Photoresist was spun onto a glass wafer and patterned using a mask.
후속연구
coll O157:H7 (9) and Salmonella typhimurium (10). However, more research is needed to improve the performance of Salmonella detection in food.
참고문헌 (12)
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