研究目的
To develop a sensitive lateral ?ow immunoassay (LFIA) for detecting Ralstonia solanacearum, the causal agent of potato brown rot, by enlarging gold nanoparticles to lower the detection limit.
研究成果
The gold enhancement approach significantly reduces the detection limit of Ralstonia solanacearum by 33 times to 3 × 10^4 cells/mL in potato tuber extracts, enabling diagnosis of latent infections. It is rapid (3 min for enhancement, 15 min total), simple, and reproducible, making it promising for field-based diagnostic applications despite some matrix effects.
研究不足
The gold enhancement method showed some nonspecific staining in potato tuber extracts due to matrix components, which affected the detection limit compared to buffer medium. The detection limit achieved (3 × 10^4 cells/mL) is higher than that of silver enhancement (200 cells/mL) and PCR-based methods, indicating room for improvement in sensitivity and matrix tolerance.
1:Experimental Design and Method Selection:
Used a sandwich LFIA format with gold nanoparticles (GNPs) as labels and post-assay gold enhancement via reduction of tetrachloroauric (III) anions with hydroxylamine to increase GNP size and signal intensity.
2:Sample Selection and Data Sources:
Ralstonia solanacearum strain NCPPB No. 2316 and other potato pathogens were used; potato tuber extracts were prepared from homogenized tubers.
3:List of Experimental Equipment and Materials:
Included GNPs synthesized by the Frens method, antibodies, membranes (e.g., CNPC-12μ nitrocellulose), enhancement solutions (HAuCl4 and hydroxylamine), and various microscopes and readers.
4:Experimental Procedures and Operational Workflow:
Synthesized GNP-antibody conjugates, fabricated test strips, performed LFIA by dipping strips in samples for 10 min, applied enhancement solution for 3 min, washed, and analyzed visually or with a scanner.
5:Data Analysis Methods:
Used scanning electron microscopy (SEM), atomic force microscopy (AFM), dynamic light scattering (DLS), transmission electron microscopy (TEM), and image analysis software (e.g., Image Tool, TotalLab TL120) for characterization and quantification.
独家科研数据包,助您复现前沿成果,加速创新突破
获取完整内容-
EnSpire plate reader
EnSpire
Perkin Elmer
Measuring optical density at 580 nm for flocculation curve determination
-
JEM-1400 microscope
JEM-1400
Jeol
Transmission electron microscopy for GNP imaging
-
Zetasizer Nano
Zetasizer Nano
Malvern Panalytical
Dynamic light scattering for hydrodynamic size measurement
-
JSM-6510LV microscope
JSM-6510LV
Jeol
Scanning electron microscopy for GNP imaging
-
Cannon 9000F Mark II scanner
Cannon 9000F Mark II
Canon
Scanning test strips for quantitative analysis
-
dialysis bag
10 × 6 mm
Sigma-Aldrich
Separating protein molecules from low-molecular-weight compounds
-
Libra S80 spectrophotometer
Libra S80
Biochrom
Measuring optical density at 520 nm for GNP characterization
-
SmartSPM-1000 atomic force microscope
SmartSPM-1000
AIST-NT
Atomic force microscopy for surface characterization
-
IsoFlow dispenser
IsoFlow
Imagene Technology
Dispensing reagents on nitrocellulose membranes
-
Index Cutter-1 automatic guillotine
Index Cutter-1
A-Point Technologies
Cutting lateral flow multimembrane composite into test strips
-
FR-900 continuous band sealer
FR-900
Dingli Packing Machinery
Packing test strips in laminated aluminum foil bags
-
IB-3 coater
IB-3
EIKO
Coating membranes with gold for SEM
-
MilliQ filter
MilliQ
Millipore
Producing deionized water for solutions
-
登录查看剩余11件设备及参数对照表
查看全部