Recently, the importance of on-site detection of pathogens has drawn attention in the field of molecular diagnostics. Unlike in a laboratory environment, on-site detection of pathogens is performed under limited resources. In this study, we tried to optimize the experimental conditions for on-site detection of pathogens using a combination of ultra-fast convection polymerase chain reaction (cPCR), which does not require regular electricity, and nucleic acid lateral flow (NALF) immunoassay. Salmonella species was used as the model pathogen. DNA was amplified within 21 minutes (equivalent to 30 cycles of polymerase chain reaction) using ultra-fast cPCR, and the amplified DNA was detected within approximately 5 minutes using NALF immunoassay with nucleic acid detection (NAD) cassettes. In order to avoid false-positive results with NAD cassettes, we reduced the primer concentration or ultra-fast cPCR run time. For singleplex ultra-fast cPCR, the primer concentration needed to be lowered to 3 μM or the run time needed to be reduced to 14 minutes. For duplex ultra-fast cPCR, 2 μM of each primer set needed to be used or the run time needed to be reduced to 14 minutes. Under the conditions optimized in this study, the combination of ultra-fast cPCR and NALF immunoassay can be applied to on-site detection of pathogens. The combination can be easily applied to the detection of oral pathogens.

Antibiotic Detection Kit (Combination I), a lateral flow immunoassay (LFIA) developed for detecting antibiotic residues in milk, was utilized for the analysis of antibiotic residues in the muscle tissue of olive flounder. After 5-h treatment of samples by placing them in water dosed with sulfadimethoxine (SDM; 200 g/ton water), the residue depletion of SDM was investigated in 25 cultured olive flounders (Paralichthys olivaceus). Muscles from fish were sampled before treatment and on the 1st, 2nd, 3rd, 4th and 5th days after treatment. The concentration of SDM in the muscle was then determined by LFIA. The absorbance ratio of the sample to the control blank (Bs/Bo) was employed as an index to determine the residue in olive flounder muscle. To investigate the recovery rate, standard solutions were added to muscle samples to obtain final concentrations of 25 and 50 ng/mL in the muscle. The recovery rates of all spiked samples were >96.6% of the spiked value. SDM was detectable in the muscle of fish treated with the drug until the 1st day of the withdrawal period. The present study shows that the LFIA can be easily adopted to detect SDM residues in the tissue of farmed fish.

Antibiotic Detection Kit (Combination I), a lateral flow immunoassay (LFIA) developed for the detection of antibiotic residues in milk, was utilized for the analysis of antibiotic residues in the muscle tissue of olive flounder. After 60-min treatment by dipping in water dosed with ampicillin (200-g/ton water), the residue depletion of ampicillin was investigated in 25 cultured olive flounder (Paralichthys olivaceus). Muscles of fish were sampled on the 1st, 2nd, 3rd, 4th and 5th day after drug treatment. The concentration of ampicillin in the muscle was determined by LFIA. The absorbance ratio of the sample to the control blank (Bs/Bo) was employed as an index to determine the muscle residues in olive flounder. To investigate the recovery rate, standard solutions were added to muscle samples to give final concentrations in the muscle of 4 and 8 ng/ml. The recovery rates of all spiked samples were > 96% of the spiked value. Ampicillin was detected in the muscle of fish treated with the drug until the 2nd day of the withdrawal period. The present study showed that the LFIA can be easily adopted to predict ampicillin residues in tissue of farmed fishes.

In this study, a rapid immuno-chromatographic assay was developed to detect Salmonella Typhimurium. This assay was based on the lateral flow immunoassay. Two different anti-Salmonella polyclonal antibody (Pab) are used in this strip assay for the double antibody sandwich immunoassay method. The strip sensor was composed of sample pad, conjugation pad, membrane, and absorbent pad. For the optimal conjugation between nano-gold and anti-Salmonella Pab, the pH value of colloidal gold solution was determined to 8.4. To prevent non-specific binding between nano gold and anti-Salmonella Pab, sample pad and membrane was treated with PBS buffer, included 2% BSA and 10% sucrose. The control and detection zones were visually detected according to antigen-antibody binding. The detection limit of the developed strip assay was 105 CFU/mL within 10 min, and improved until 104 CFU/mL after 12 hr.

This study was performed to develop and optimize a rapid strip kit for the dual-detection of Aflatoxin B1 (AF) and Ochratoxin A (OTA). The strip kit is composed of sample pad, conjugation pad, nitrocellulose membrane, and absorbent pad. Manually spotted mycotoxin-BSA conjugates and anti-mouse IgG on nitrocellulose membrane were used as the test line and the control line, respectively. Conditions for rapid and easy detection of mycotoxins, conjugation between toxin-MAbs and nano Au particles, pretreatment method of pads, and composition of standards solutions were optimized. Feasibility of the strip kit to detect AF and OTA were evaluated with standard samples. Test results were acquired in 10 min and the detection limits were 100 ppm for AF and OTA, respectively.

최근 세계 곳곳에서 강우량증가에 의한 홍수피해가 속출하고 있다. 돌발적으로 집중되는 강우에 의해 하천이 범람하고 제방이 붕괴되는 피해가 증가하는 추세이다. 하도 내 홍수위가 증가하여 제방이 월류하면 제방은 그 기능을 상실하여 점진적인 붕괴가 진행된다. 제방의 점진적인 붕괴는 많은 연구가 진행중이나 복잡한 붕괴 메커니즘과 다양한 영향인자들로 인해 이론적으로 명확게 규명되지 않았다. 또한 대부분의 기존연구들이 제방의 붕괴를 댐붕괴와 동일하게 가정하거나 급격한 붕괴 또는 하도의 흐름에 횡방향으로 분류가되는 횡월류 위어와 동일하게 가정하여 실제 제방붕괴 양상과는 상이측면이 있다. 따라서 본 연구에서는 비점착성 재질로 제방을 축조하고 하도의 흐름이 존재할 때, 하도의 횡방향으로 제방이 월류하는 수리실험을 수행하여 하도의 Froude 수와 제방의 붕괴폭 및 붕괴유량과의 관계를 분석하였다. 그 결과 제방의 붕괴유량은 횡월류 위어와는 다르게 하도의 흐름과 제방 단면의 충돌에 의한 2차적인 영향이 발생하였고 이로 인해 붕괴유량의 차이가 발생하였다. 또한 붕괴유량이 수평한 제내지를 전파하는 실험을 수행하여 하도조건에 따른 붕괴홍수파 전파각의 시간적인 변화를 분석하였다. 붕괴홍수파 전파각은 붕괴의 발달 정도에 따라 상류 또는 하류전파각이 변화하는 것을 확인하였다. 본 연구 결과는 댐붕괴나 횡월류 위어와는 구분되는 제방의 붕괴특성을 규명하였으며 이러한 특성이 제방붕괴에 대한 비상대처계획 수립 시에 적용한다면 최종 침수면적이나 침수심을 고려한 공간적인 계획뿐만 아니라 붕괴양상에 따른 시간적인 인자를 함께 고려한 계획이 수립될 수 있을 것으로 기대된다.