• Korean Journal of Veterinary Research
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• v.13 no.1
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• pp.39-46
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• 1973

Cultural method, dark field microscopy & fluorescent antibody technique were compared for their sensitivity of the detection of leptospires from experimentally infected mice. Two groups of mice were infected with L. icterohemorrhagiae (M20) and L. australis (Ballico), and the infected blood, urine and a number of organs were subjected to the bacterial isolation. The results obtained were summarized as follows: 1. L. icterohemorrhagiae (M20) and L. australis (Ballico) in blood, urine and various tissues of experimentally infected mice were detected with a negrigible non specificity, by the fluorescent antibody technique. 2. The fluorescent antibody technique, as applied to detection of leptospires in blood, urine and various infected tissue, proved to be better than cultural method and dark-field microscopy. 3. Early detection of leptospires by fluorescent antibody technique were possible in blood at 2 days after inoculation, whereas detection of organisms in liver, spleen, lung and kidney were observed later. By means of fluorescent antibody technique, the detection of leptospires in kidney and urine was possible up to 34 days postinoculation, whereas those in other parts were impossible. 4. Fluorescent antibody reaction of leptospires were highly specific to homologous antigen rather than to heterologous one. 5. Fluorescent antibody technique may be of value in the application for the demonstration of leptospira from clinical specimens.

• Korean Journal of Veterinary Research
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• v.14 no.2
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• pp.243-252
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• 1974

The experiment was performed in order to investigate the applicability of the rapid detection of salmonellae in various animal-origin foods by means of the direct fluorescent antibody technique. Egg, sausage and chicken were inoculated with various concentrations of Sal.paratuphi A, Sal. paratyhi B and Sal. thompson, and the fluorescent antibody technique was applied and compared with the conventional cultura method for the sensitivity of detection of the organisms. Two methods were employed in the fluorescent antibody technique; the direct smear method in which the smear being made directly from the specimens, and the enrichment smear method in which the smear being made from the enrichment broth. The effect of various enrichment time (1,5,8,11 and 13 hours) in tetrathionate broth on the detection of salmonellae in the fluoresent antibody technique was also studied. The results obtained were summarized as followings; 1. Of the three methods, the enrichment smear method of fluorescedt antibody technique was highly effective as cultural method for the detection of salmonella organisms. 2. Direct smear method of fluorescent antibody technique was effective as two other methods $5{\times}10^4$ organisms presented in 50 g(ml) of specimens. This method may not be applicable when the specimens contained $5{\times}10^2$ or less organisms. 3. Of the three specimens, the recovery rate of Salmonella organisms from egg was slightly higher than that of sausage and chicken. 4. In fluorescent antibody technique and cultural method, the specimens inoculated with Sal. thompson were found to be higher detection rate than the specimens inoculated with Sal. paratyphi A, 5. The optimum enrichment time of Salmonella organisms in tetrathionate broth on the detection by fluorscent antibody technique was found to be 11 hours or longer when the specimens of egg, sausage and chicken were inoculated with approximately 500 organisms. The longer enrichment time was the higher detection rate up to 11 hours tested.

• Fisheries and Aquatic Sciences
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• v.4 no.1
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• pp.32-38
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• 2001

Several molecular biological techniques have been used to detect virus rapidly and accurately, but these methods have limitations in the early stage of viral infection with very low concentration of virus. We compared the detection limits of IMS-PCR, Western blot and ELISA with infectious hematopoietic necrosis virus OHNV). Four antibodies, rabbit anti-IHNV polyclonal antibody, anti-IHNV nucleocapsid protein monoclonal antibody, anti-IHNV nucleocapsid protein polyclonal antibody, and anti-IHNV glycoprotein polyclonal antibody, were tested to find out the most effective antibody for each method. The detection limit with IMS- PCR was $2\times10^6$ pfu when the viral RNA was extracted before RT-PCR. In the western blot with rabbit anti­IHNV polyclonal antibody one pfu of virus could be detected. In ELISA, 10 pfu of virus particles were detected with the same antibody.

• Bulletin of the Korean Chemical Society
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• v.27 no.4
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• pp.479-483
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• 2006

We developed a microfluidic immunoassay platform for the detection of various analytes such as bacterial pathogen by packing antibody-immobilized glass beads in spatially-isolated microchambers on a microfluidic device. Primary amines of antibody were covalently conjugated to carboxyl-terminated glass beads previously treated with aminosilane followed by glutaraldehyde. Through this covalent binding, up to 905 $\mu$g immunoglobulin G (IgG) per gram of glass beads was immobilized. For application, glass beads attaching antibody specific to Escherichia coli O157:H7, a foodborne pathogen, were packed into a microfluidic device and used for the detection of the serotype. This prototype immunoassay device can be used for the simultaneous detection of multiple analytes by sequentially packing different-sized glass beads attaching different antibody in discrete microchambers on a single microfluidic device.

• Parasites, Hosts and Diseases
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• v.47 no.2
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• pp.153-157
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• 2009

Diagnosis of hydatidosis is based on immunodiagnostic methods along with radiological and ultrasound examinations. The objectives of the present study were to develop a specific and simple antigen-based ELISA method for diagnosis of hydatidosis and compare it with antibody detection method. The subjects in this study included 89 patients in the following groups: surgically confirmed hydatidosis patients (35 cases), control with other parasitic diseases (29 cases), and healthy controls (25 cases). Hyperimmune serum was raised against hydatid cyst fluid in rabbits. Anti-hydatid cyst IgG was purified by affinity chromatography using protein A column and labeled with horseradish peroxidase. Collected sera were assessed for hydatid cyst antigens and antibody by ELISA. Circulating hydatid antigen was found in 9 out of 35 patients with surgically confirmed hydatidosis. A sensitivity of 25.7% and a specificity of 98.0% were calculated for the antigen detection assay. Antibody detection by indirect ELISA, using antigen B, showed that 94.2% of patients (33 cases) have anti-hydatid cyst antibodies in their serum while cross reaction was noted in a few of non-hydatidosis patients. A sensitivity of 94.2% and specificity of 81.6% were found for the antibody detection assay. Findings of this study indicated that antibody detection assay is a sensitive approach for diagnosis of hydatid cyst while antigen detection assay might be a useful approach for assessment of the efficacy of treatment especially after removal of the cyst.

• Journal of Information Processing Systems
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• v.15 no.1
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• pp.137-150
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• 2019

Although the research of immune-based anomaly detection technology has made some progress, there are still some defects which have not been solved, such as the loophole problem which leads to low detection rate and high false alarm rate, the exponential relationship between training cost of mature detectors and size of self-antigens. This paper proposed an intrusion detection method based on changes of antibody concentration in immune response to improve and solve existing problems of immune based anomaly detection technology. The method introduces blood relative and blood family to classify antibodies and antigens and simulate correlations between antibodies and antigens. Then, the method establishes dynamic evolution models of antigens and antibodies in intrusion detection. In addition, the method determines concentration changes of antibodies in the immune system drawing the experience of cloud model, and divides the risk levels to guide immune responses. Experimental results show that the method has better detection performance and adaptability than traditional methods.

• Journal of Sensor Science and Technology
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• v.18 no.2
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• pp.160-167
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• 2009

In this study, secondary antibody-containing quartz crystal microbalance(QCM) sensor chip was prepared and utilized for the detection of human and bovine haptoglobin. Anti-goat immunoglobulin G antibody, which is a secondary antibody capable of capturing primary antibodies raised in goat, was immobilized through the reaction between hydrazide and aldehyde group prepared on the QCM surface and antibody respectively. The resulting sensor chip showed higher stability in the repeated surface regeneration with acidic dissociation solution as well as requiring lower amount of primary antibody when compared to the protein G sensor chip. The secondary antibody sensor chip was applied for the estimation of bovine and human haptoglobin.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.3
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• pp.874-879
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• 2004

Alakaline phosphatase (ALP)- or horseradish peroxidase (HRP)-antibody conjugate was used frequently on the immunological detection methods such as enzyme-linked immunosobent assay (ELISA), immunobolt, immunohistochemistry. The classical enzyme-antibody coupling method by one-step (direction) injection of glutaraldehyde bring into being disadvantage such as low sensitivity of antigen detection because of homopolymers. This study was modified with the dialysis glutaraldehyde method to provide simple coupling through E-amino residues present in most protein. The dialysis glutaraldehyde coupling effects were better than the classical one-step glutaraldehyde injection in antigen detection of ELISA and immunobolt. Optimal dose of the dialysis glutaraldehyde solution was 0.10-0.25 %. This results suggest that the dialysis glutaraldehyde coupling method can readily applied to antigen detection of in vitro and in vivo.

• The Plant Pathology Journal
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• v.25 no.4
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• pp.328-332
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• 2009

Diseases caused by Pepper yellow leaf curl virus infection is considered to be emerging plant diseases in Indonesia in the last five years. One key factor for disease management is the availability of accurate detection of the virus in plants. Polyclonal antibody for Pepper yellow leaf curl Indonesia virus-Bogor (PYLCIV-Bgr) was produced for detection of the virus using I-ELISA and DIBA methods. The antibody was able to detect PYLCIV-Bgr from infected plants up to dilution 1/16,384 and cross reaction was not observed with Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV), and Chilli veinal mottle virus (ChiVMV). Positive reaction was readily detected in membrane containing Begomovirus samples from Yogyakarta (Kaliurang and Kulonprogo) and West Java (Bogor and Segunung). Infection of PYLCIV-Bgr in chillipepper, tomato, and Ageratum conyzoides was also confirmed using polyclonal antibody for PYLCIV-Bgr in DIBA. Polyclonal antibody for PYLCIV-Bgr is suggested to be included in disease management approach due to its good detection level.

• Microbiology and Biotechnology Letters
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• v.25 no.4
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• pp.414-419
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• 1997

In order to develop an enzyme-linked immunosorbent assay (ELISA) for deoxynivalenol (DON) in com, we produced a specific monocl- onal antibody and established ELISA conditions. After the spleen cells from mice immunized with DON-bovine serum albumin conjugate were fused with S$_{p}$2/0 myeloma cells, a hybridoma cell 3G7 producing anti-DON antibody was screened by ELISA. From the standard curve of competitive direct ELISA (cdELISA) using 3G7 monoclonal antibody and DON-HRP conjugate, the detection range of DON showed 3-3,000 ng/ml (ppb). The monoclonal antibody showed some cross-reactivities against DON analogues such as 15 acetyl-DON (110%), nivalenol (5.0%), 3 acetyl-DON (1.7%), fusarenon-x (0.72%), and T-2 (0.59%). When the cdELISA was applied to the spiked coms after extracting with 60% methanol and diluting 5- fold with washing buffer, the assay recoveries of DON were 313, 163, 106, and 88.9% (av., 168%) in the levels of 200, 600, 2,000, and 6,000ng/g, respectively. For the quantitation of DON in coms, 30 samples kept under two different storage conditions of cold and room temperature were assayed by cdELISA. The mean detection concentrations were 595 (detection range, 0-2,750) and 2,448 (detection range, 0-4,500) ppb, respectively.