Abstract
A direct, sensitive, near-real-time, handheld optical immunoassay device was developed to detect Salmonella typhimurium in the naturally occurring liquid from fresh poultry packages (hereafter "chicken matrix"), with just single pipetting of sample (i.e., no filtration, culturing and/or isolation, thus reducing the assay time and the error associated with them). Carboxylated, polystyrene microparticles were covalently conjugated with anti-Salmonella, and the immunoagglutination due to the presence of Salmonella was detected by reading the Mie scatter signals from the microfluidic channels using a handheld device. The presence of chicken matrix did not affect the light scatter signal, since the optical parameters (particle size d, wavelength of incident light λ and scatter angle θ) were optimized to minimize the effect of sample matrix (animal tissues and blood proteins, etc.). The sample was loaded into a microfluidic chip that was split into two channels, one pre-loaded with vacuum-dried, antibody-conjugated particles and the other with vacuum-dried, bovine serum albumin-conjugated particles. This eliminated the need for a separate negative control, effectively minimizing chip-to-chip and sample-to-sample variations. Particles and the sample were diffused in-channel through chemical agitation by Tween 80, also vacuum-dried within the microchannels. Sequential mixing of the sample to the reagents under a strict laminar flow condition synergistically improved the reproducibility and linearity of the assay. In addition, dried particles were shown to successfully detect lower Salmonella concentrations for up to 8 weeks. The handheld device contains simplified circuitry eliminating unnecessary adjustment stages, providing a stable signal, thus maximizing sensitivity. Total assay time was 10min, and the detection limit 10CFUmL-1 was observed in all matrices, demonstrating the suitability of this device for field assays.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 342-349 |
| Number of pages | 8 |
| Journal | Biosensors and Bioelectronics |
| Volume | 40 |
| Issue number | 1 |
| DOIs | |
| State | Published - Feb 15 2013 |
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Keywords
- Chicken carcass
- Immunoagglutination
- Lab on a chip
- Latex agglutination test
- Microfluidic device
- Mie scatter
- Salmonella typhimurium
ASJC Scopus subject areas
- Biophysics
- Biomedical Engineering
- Biotechnology
- Electrochemistry
Cite this
Single-pipetting microfluidic assay device for rapid detection of Salmonella from poultry package. / Fronczek, Christopher F.; You, David J.; Yoon, Jeong-Yeol.
In: Biosensors and Bioelectronics, Vol. 40, No. 1, 15.02.2013, p. 342-349.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Single-pipetting microfluidic assay device for rapid detection of Salmonella from poultry package
AU - Fronczek, Christopher F.
AU - You, David J.
AU - Yoon, Jeong-Yeol
PY - 2013/2/15
Y1 - 2013/2/15
N2 - A direct, sensitive, near-real-time, handheld optical immunoassay device was developed to detect Salmonella typhimurium in the naturally occurring liquid from fresh poultry packages (hereafter "chicken matrix"), with just single pipetting of sample (i.e., no filtration, culturing and/or isolation, thus reducing the assay time and the error associated with them). Carboxylated, polystyrene microparticles were covalently conjugated with anti-Salmonella, and the immunoagglutination due to the presence of Salmonella was detected by reading the Mie scatter signals from the microfluidic channels using a handheld device. The presence of chicken matrix did not affect the light scatter signal, since the optical parameters (particle size d, wavelength of incident light λ and scatter angle θ) were optimized to minimize the effect of sample matrix (animal tissues and blood proteins, etc.). The sample was loaded into a microfluidic chip that was split into two channels, one pre-loaded with vacuum-dried, antibody-conjugated particles and the other with vacuum-dried, bovine serum albumin-conjugated particles. This eliminated the need for a separate negative control, effectively minimizing chip-to-chip and sample-to-sample variations. Particles and the sample were diffused in-channel through chemical agitation by Tween 80, also vacuum-dried within the microchannels. Sequential mixing of the sample to the reagents under a strict laminar flow condition synergistically improved the reproducibility and linearity of the assay. In addition, dried particles were shown to successfully detect lower Salmonella concentrations for up to 8 weeks. The handheld device contains simplified circuitry eliminating unnecessary adjustment stages, providing a stable signal, thus maximizing sensitivity. Total assay time was 10min, and the detection limit 10CFUmL-1 was observed in all matrices, demonstrating the suitability of this device for field assays.
AB - A direct, sensitive, near-real-time, handheld optical immunoassay device was developed to detect Salmonella typhimurium in the naturally occurring liquid from fresh poultry packages (hereafter "chicken matrix"), with just single pipetting of sample (i.e., no filtration, culturing and/or isolation, thus reducing the assay time and the error associated with them). Carboxylated, polystyrene microparticles were covalently conjugated with anti-Salmonella, and the immunoagglutination due to the presence of Salmonella was detected by reading the Mie scatter signals from the microfluidic channels using a handheld device. The presence of chicken matrix did not affect the light scatter signal, since the optical parameters (particle size d, wavelength of incident light λ and scatter angle θ) were optimized to minimize the effect of sample matrix (animal tissues and blood proteins, etc.). The sample was loaded into a microfluidic chip that was split into two channels, one pre-loaded with vacuum-dried, antibody-conjugated particles and the other with vacuum-dried, bovine serum albumin-conjugated particles. This eliminated the need for a separate negative control, effectively minimizing chip-to-chip and sample-to-sample variations. Particles and the sample were diffused in-channel through chemical agitation by Tween 80, also vacuum-dried within the microchannels. Sequential mixing of the sample to the reagents under a strict laminar flow condition synergistically improved the reproducibility and linearity of the assay. In addition, dried particles were shown to successfully detect lower Salmonella concentrations for up to 8 weeks. The handheld device contains simplified circuitry eliminating unnecessary adjustment stages, providing a stable signal, thus maximizing sensitivity. Total assay time was 10min, and the detection limit 10CFUmL-1 was observed in all matrices, demonstrating the suitability of this device for field assays.
KW - Chicken carcass
KW - Immunoagglutination
KW - Lab on a chip
KW - Latex agglutination test
KW - Microfluidic device
KW - Mie scatter
KW - Salmonella typhimurium
UR - http://www.scopus.com/inward/record.url?scp=84868631352&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84868631352&partnerID=8YFLogxK
U2 - 10.1016/j.bios.2012.07.076
DO - 10.1016/j.bios.2012.07.076
M3 - Article
VL - 40
SP - 342
EP - 349
JO - Biosensors
JF - Biosensors
SN - 0956-5663
IS - 1
ER -