• Korean Journal of Microbiology
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• v.39 no.2
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• pp.114-117
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• 2003

In molecular biology, it is necessary to develop an easy and rapid method to identify a specific DNA sequence. Though Southern and Northern blot techniques have been used widely for the analysis of gene structure and function, those methods are inconvenient in the points that we need to control incubation temperature, time, and other parameters to get the final result. In this study, we report a new method for the rapid analysis of specific DNA sequence with the modification of an immunochromatographic method. The lateral flow DNA analysis strip is composed of a sample pad, a nitrocellulose membrane for the separation and propagation of analytes, and an absorption pad for the generation of capillary action. Capture DNA was immobilized on the membrane by UV cross-linking and target DNA was labeled with Cy-5 for signaling. The samples containing target DNA were applied onto the sample pad, incubated for 15 min for separation, and scanned with a GSI fluorescence scanner. Though the hybridization reaction occurs in a short time without any washing steps, there appears to be little cross hybridization between the different sequences. The result showed a possibility that the new method can be used for the rapid identification of specific DNA sequence among the samples.

• Journal of Biosystems Engineering
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• v.36 no.2
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• pp.140-146
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• 2011

This study was performed to develop a rapid test kit for pathogenic Salmonella in various samples. The rapid detection kit has been fabricated based on nitrocellulose lateral-flow strip. Colloidal gold and biotin conjugated Salmonella antibodies were used as a tag and a receptor, respectively. Manually spotted Salmonella antibody and Neutravidin on nitrocellulose membrane were used as test and control lines, respectively. Feasibility of the rapid kit to detect Salmonella typhimurium in samples were evaluated. The intensity of the color of the test line started to increase with the samples in which higher concentration of the cells were contained. The sensitivity of the sensor was $10^6$ cfu/mL Salmonella spiked in PBS. Also, the rapid test kit could detect $10^6$ cfu/mL of Salmonella in chicken meat extract.

• Korean Journal of Agricultural Science
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• v.40 no.2
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• pp.139-146
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• 2013

This study was performed to develop a rapid test kit for pathogenic Staphylococcus in various samples. The rapid detection kit has been fabricated based on nitrocellulose lateral-flow strip. Colloidal gold and Staphylococcus antibodies were used as a tag and a receptor, respectively. Manually spotted Staphylococcus antibody and anti-mouse antibody on the surface of nitrocellulose membrane were used as test and control lines, respectively. Feasibility of the rapid kit to detect Staphylococcus aureus in samples were evaluated. The intensity of the color of the test line started to increase with the samples in which higher concentration of the cells were contained. The sensitivity of the sensor was $10^6$ cfu/mL Staphylococcus spiked in PBS. Also, the rapid test kit could detect $10^5$ cfu/mL of Staphylococcus in chicken meat extract.

• Korean Journal of Agricultural Science
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• v.39 no.4
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• pp.613-618
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• 2012

This study was performed to optimize the basic platform of a lateral flow immunoassay. Improvement of the limit of detection (LOD) was evaluated according to the width of a nitrocellulose membrane with varying concentrations of analyte. The analyte, neutravidin was detected based on the avidin-biotin interaction. The antibody-Au nanoparticle conjugation was mostly stabled in a PBS buffer of pH 7.3. The optimal widths of a nitrocellulose membrane were 4 and 6 mm considering the sample flow rate and signal strength of the test line on the membrane. The LOD of neutravidin was 0.001 mg/ml in the optimum conditions.

• Applied Chemistry for Engineering
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• v.33 no.2
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• pp.173-178
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• 2022

A lateral flow immunoassay (LFIA) method using carbon nanodot@silica as a signaling material was developed for analyzing the concentration of retinol-binding protein 4 (RBP4), one of the lung cancer biomarkers. Instead of antibodies mainly used as bioreceptors in nitrocellulose membranes in LFIA for protein detection, aptamers that are more economical, easy to store for a long time, and have strong affinities toward specific target proteins were used. A 5' terminal of biotin-modified aptamer specific to RBP4 was first reacted with neutravidin followed by spraying the mixture on the membrane in order to immobilize the aptamer in a porous membrane by the strong binding affinity between biotin and neutravidin. Carbon nanodot@silica nanoparticles with blue fluorescent signal covalently conjugated to the RBP4 antibody, and RBP4 were injected in a lateral flow manner on to the surface bound aptamer to form a sandwich complex. Surfactant concentrations, ionic strength, and additional blocking reagents were added to the running buffer solution to optimize the fluorescent signal off from the sandwich complex which was correlated to the concentration of RBP4. A 10 mM Tris (pH 7.4) running buffer containing 150 mM NaCl and 0.05% Tween-20 with 0.6 M ethanolamine as a blocking agent showed the optimum assay condition for carbon nanodot@silica-based LFIA. The results indicate that an aptamer, more economical and easier to store for a long time can be used as an alternative immobilizing probe for antibody in a LFIA device which can be used as a point-of-care diagnosis kit for lung cancer diseases.

• Korean Journal of Agricultural Science
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• v.39 no.3
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• pp.421-425
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• 2012

The objective of this study was to improve the detection limit of rapid detection kit for Salmonella Typhimurium by image analysis system. The rapid detection kit was comprised of four elements: sample pad, conjugate pad, nitrocellulose pad and absorbent pad. Gold nanoparticle and Salmonella antibody were used as a tag and a receptor. Salmonella antibody and goat rabbit IgG antibody were used as test and control lines on nitrocellulose membrane. The color intensity of test line began to increase from $10^5CFU/mL$ of Salmonella sample. A multiple linear regression analysis was employed to explain the relationship between predicted and measured number of Salmonella cells. The developed model could successfully predict the cell number of Salmonella with validation against extra-experimental result.

• Bulletin of the Korean Chemical Society
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• v.30 no.12
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• pp.2999-3005
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• 2009

Among the more than 120 different types of human papillomavirus (HPV), types 16 and 18 have been known to be high risk agents that cause cervical cancer. We examined, in an immuno-chromatographic analysis, the potential of using the early gene product, E7 protein, as a diagnostic marker of cervical cancer caused by HPV. We developed monoclonal antibodies specific to HPV-16 and 18 E7 proteins that were produced from bacterial cells using gene recombinant technology. For each E7 protein, the optimal antibody pair was selected using the immuno-chromatographic sandwichtype binding system based on the lateral flow through membrane pores. Under these conditions, this rapid testing assay had a detection capability as low as 2 ng/mL of E7 protein. Furthermore, since viral analysis required the host cell to be lysed using chemicals such as detergents, it was possible that the E7 protein was structurally damaged during this process, which would result in a decrease in detection sensitivity. Therefore, we examined the detrimental effects caused by different detergents on the E7 protein using HeLa cells as the host. In these experiments, we found that the damage caused by the detergent, nonylphenylpolyethylene glycol (NP-40), was minimal relative to Triton X-100 commonly used for the cell lysis. Temperature also affected the stability of the E7 protein, and we found that the E7 protein was stabilized at 4$^{\circ}C$ for about 2 h, which was 4 times longer than at room temperature. Finally, a HPV-infected cervical cancer cell line, which was used as a real sample model, was treated using the optimized conditions and the presence of E7 proteins were analyzed by immuno-chromatography. The results of this experiment demonstrated that this rapid test could specifically detect HPV-infected samples.