• Parasites, Hosts and Diseases
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• v.41 no.4
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• pp.203-207
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• 2003

An ELISA was developed for the diagnosis of vivax malaria using multiple stage-specific recombinant antigens of Plasmodium vivax. The DNA from the whole blood of a malaria patient was used as template to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1). Each amplified DNA fragment was inserted into pQE30 plasmid to induce the expression of His-tagged protein in Escherichia coli (M15 strain) by IPTG. His-tagged proteins were purified by Ni-NTA metal-affinity chromatography and used as antigens for ELISA with patient sera that were confirmed previously by blood smear examinations. When applied to patient sera, 122 (80.3%) out of 152 vivax malaria cases reacted to at least one antigen, while no reactions were observed with 128 uninfected serum samples. We applied this ELISA to the screening of 3,262 civilian residents in endemic regions near the DMZ, which resulted in 236 positively detected (7.2%) cases. This method can be applied to serological diagnosis and mass screening in endemic regions, or can be used as a safety test for transfusion blood in endemic areas.

• Korean Journal of Clinical Laboratory Science
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• v.43 no.1
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• pp.1-5
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• 2011

The infection rate of syphilis is still increasing in the world especially in developing countries and the infection is often seen in large amounts of clinical specimens. For the diagnosis of this disease, Rapid Plasma Reagin (RPR)/Venereal Disease Research Laboratory (VDRL) has still been used as one of major primary methods to diagnose syphilis even though the test readings are somewhat subjective with high false positive rates. Recently, the automatic ARCHITECT Syphilis TP, which is based on the detection of the TP-specific antibodies, has been introduced in many laboratories. Therefore, the clinical assessment of the method is needed to provide primary diagnosis of syphilis at the moment. We evaluated 3 different manual rapid kits and ARCHITECT Syphilis TP comparing with RPR/FTA-ABS and analysed their diagnostic properties. From February 2006 to April 2008, 203 positive and 250 negative specimens, obtained from Chungbuk National University Hospital were used for the evaluation. In the evaluation between manual rapid kits, their specificities were as high as 99.2 ~ 99.6% while their sensitivities were observed with little differences; 98.0% (199/203) for Kit A, 96.6% (196/203) for Kit B, and 97.4% (197/203) for Kit S. In the case of ARCHITECT Syphilis TP test, it showed 100% specificity (250/250) and 98.5% sensitivity (249/250). Kappa values comparing with RPR/FTA-ABS were 0.978 for Kit A, 0.964 for Kit B and Kit S, and 0.987 for ARCHITECT Syphilis TP. From our evaluation, we found out that manual rapid tests and ARCHITECT Syphilis TP have very good clinical accuracies and high kappa agreements with RPR/FTA-ABS. Due to its automation and quick simultaneous diagnosis with another serological markers, we suggest that the ARCHITECT Syphilis TP is one of best suitable method for the primary diagnosis of syphilis and that it might be able to replace RPR method in the laboratories.

• Journal of Food Hygiene and Safety
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• v.17 no.3
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• pp.152-156
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• 2002

Totally, 742 fishery products were collected at Garak wholesale market in Seoul from Jan. to Dec. in 2001. Vibrio parahaemolyticus were 12.5%(93/742). The detection ratio of V. parahaemolyticus by month was reached the maximum in summer season (July to September). The highest ratio with 22.9% were abtained from molluscs, followed by shellfishes with 15.3%, and fishes and crustaceans with about 8%. The ratio of serotypable strains were 15.2%(10/66) and identified serotyps of V.parahaemolyticus were K24, K28, K34, K42, K47 and K53

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.12
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• pp.5265-5272
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• 2016

Objective: The present study was conducted to evaluate invasive and noninvasive diagnostic methods for detection of Helicobacter pylori (H. pylori) in patients admitted with dyspeptic complaints and to compare sensitivities and specificities. Method: Sets of four gastric biopsy specimens were obtained from a total of 126 patients included in the study. The presence of H. pylori was determined by invasive tests including culture, rapid urease test, polymerase chain reaction (PCR) and histopathology. Among noninvasive tests, urea breath test, serological tests and enzyme-linked immunosorbent assay (ELISA) were performed. Results: H. pylori was isolated in 79 (62.7%) gastric biopsy cultures, whereas positivity was concluded for 105 (83.3%) patients by rapid urease test, for 106 (84.1%) by PCR, for 110 (87.3%) by histopathology, for 119 (94.4%) by urea breath test, and for 107 (84.9%) by ELISA. In the present study, the culture findings and histopathological examination findings were accepted as gold standard. According to the gold standard, urea breath test had the highest sensitivity (96.5%) and the lowest specificity (30%), whereas culture and histopathology had the highest specificities (100%). Conclusion: The use of PCR invasively with gastric biopsy samples yielded parallel results with the gold standard. PCR can be recommended for routine use in the diagnosis of H. pylori.

• Journal of Microbiology and Biotechnology
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• v.31 no.4
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• pp.520-528
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• 2021

Plesiomonas shigelloides, a member of the family Vibrionaceae, is a gram-negative, rod-shaped, facultative anaerobic bacterium with flagella. P. shigelloides has been isolated from such sources as freshwater, surface water, and many wild and domestic animals. P. shigelloides contains 102 O-antigens and 51 H-antigens. The diversity of O-antigen gene clusters is relatively poorly understood. In addition to O1 and O17 reported by other laboratories, and the 12 O serogroups (O2, O10, O12, O23, O25, O26, O32, O33, O34, O66, O75, and O76) reported previously by us, in the present study, nine new P. shigelloides serogroups (O8, O17, O18, O37, O38, O39, O44, O45, and O61) were sequenced and annotated. The genes for the O-antigens of these nine groups are clustered together in the chromosome between rep and aqpZ. Only O38 possesses the wzm and wzt genes for the synthesis and translocation of O-antigens via the ATP-binding cassette (ABC) transporter pathway; the other eight use the Wzx/Wzy pathway. Phylogenetic analysis using wzx and wzy showed that both genes are diversified. Among the nine new P. shigelloides serogroups, eight use wzx/wzy genes as targets. In addition, we developed an O-antigen-specific PCR assay to detect these nine distinct serogroups with no cross reactions among them.

• ANNALS OF ANIMAL RESOURCE SCIENCES
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• v.28 no.2
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• pp.78-96
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• 2017

Foot-and-mouth disease (FMD) is a infection that can easily spread when it occurs and causes serious economic damage because of the existence of multiple serotypes of the virus and extreme contagiousness. The most effective method in preventing the transmission of FMD virus (FMDV) is the culling of livestock and additional vaccination in the other areas depending on the spreading rate and situation. Diagnostic methods are utilized not only for the definite diagnosis of FMD but also for identification of serotype, and confirmation of antibody production after vaccination. Although many methods have been developed to diagnose, they are not still enough to detect accurately the disease in a short time. Therefore, it has been needed new diagnostic methods improved from existing methods. Previous methods were based on the enzyme-linked immunosorbent assay (ELISA) as a serological diagnostic method, or polymerase chain reaction (PCR), which is a molecular genetic method. The recent technology has been performing about the combination of both methods and how to make it faster, less costly, more sensitive and accurate way.

• Journal of Veterinary Science
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• v.21 no.4
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• pp.63.1-63.9
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• 2020

Background: Canine adenovirus type 2 (CAV-2) induces infectious laryngotracheitis in members of the family Canidae, including dogs. To date, no ELISA kits specific for CAV-2 antibody have been commercialized for dogs in Korea. Objectives: We aimed to develop new indirect enzyme-linked immunosorbent assay (I-ELISA) to perform rapid, accurate serological surveys of CAV-2 in dog serum samples. Methods: In total, 165 serum samples were collected from dogs residing in Chungbuk and Gyeongbuk provinces between 2016 and 2018. The Korean CAV-2, named the APQA1701-40P strain, was propagated in Madin-Darby canine kidney cells and purified in an anion-exchange chromatography column for use as an antigen for I-ELISA. The virus-neutralizing antibody titers of CAV-2 in the dog sera were measured by virus neutralization (VN) test. Results: We compared the results obtained between the VN and new I-ELISA tests. The sensitivity, specificity, and accuracy of new I-ELISA were 98.6%, 86.4% and 97.0% compared with VN test, respectively. New I-ELISA was significantly correlated with VN (r = 0.91). Conclusions: These results indicate that new I-ELISA is useful for sero-surveillance of CAV-2 in dog serum.

• Journal of Food Hygiene and Safety
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• v.18 no.3
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• pp.118-124
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• 2003

For the rapid detection of various pathogenic microorganisms from food sample, various kinds of kits have been developed and commercially available in the markets. With the advantages of speed, accuracy and easiness, the market of these kits has gradually increased for the QC and QA field of food company as well as testing facilities or laboratories. In this study, the characteristics such as the detection limit and the sensitivity of immunochromatographic type of rapid detection kit (Donga Co, Korea, D-kit) for E. coli 0157:H7 developed by monoclonal antibody were examined and also the possibility of application of the kit to food samples was evaluated. The reference kits used for comparison study were Reveal E. coli 0157:H7 (Neogen Co., USA, R-kit) and VIP EHEC kit (Biocontrol Inc., USA, V-kit) occupying major market share. In the detection limit test with the E. coli 0157:H7 reference, both R-kit and D-kit showed a distinct positive reaction in $10^4$/ml and weak positive reaction in $10^3$/ml, whereas V-kit showed a same reaction in 105/ml. Also, it was identified that the culture treated with heat showed more sensitivity than no heat treated culture. The sensitivity test was conducted against 22 isolates of E. coli 0157:H7, 7 strains of non-O157:H7 verotoxin-producing E. coli, 40 strains of E. coli with different O and H antigen type, and 38 strains of non-E. coli Enterobacteriaceae, and all of the test strains except three were showed exactly three were showed exactly the same reaction against three kinds of the tested kits. All the three kinds of kits showed a positive reaction against E. coli O157:H19, E. coli O148:H18 and Salmonella galinarium. We suppose that there might be a similarity in serological property between these three strains and O157:H7. From the test results, it can be concluded that there is (was) no difference between the D-kit developed in this study and R-kit or V-kit based on the detection limit and sensitivity.

• Journal of the Korean Society for Nondestructive Testing
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• v.31 no.5
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• pp.487-493
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• 2011

Alpha fetoprotein(AFP), which is serological marker for hepatocellular carcinoma, was quantitatively measured by its normal concentration, 10 ng/ml, with a label-free piezoelectric microcantilever mass sensor. The principle of detection is based on changes in the resonant frequency of the piezoelectric microcantilever before and after target molecules are attached to it, and its resonant frequency is measured electrically using a conductance spectrum. The resonant frequency of the developed sensor is approximately 1.34 MHz and the mass sensitivity is approximately 175 Hz/pg. The sensor has high reliability as mass sensor by reducing the effect of surface stress on resonant frequency due to attached proteins. 'Dip and dry' technique was used to react the sensor with reagents for immobilizing AFP antibody on the sensor and detecting AFP antigen. The measured mass of the detected AFP antigen was 6.02 pg at the concentration of 10 ng/ml, and 10.67 pg at 50 ng/ml when the immunoreaction time was 10 min.

• Biomedical Science Letters
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• v.3 no.2
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• pp.95-105
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• 1997

Currently, the laboratory diagnosis for lyme disease have been performed with the detection of antibodies against Borrelia burgdorferi. However there might be some difficulties in the interpretation of obtained results due to the usage of foreign isolates as an antigen in the test. Therefore the optimization of serological tests with Korean isolates of B. burgdorferi as an antigen would be needed to establish the standardized diagnostic method for the detection of antibodies against B. burgdorferi infection. In this study, the optimization of ELISA was investigated with experimentally challenged rabbit sera and sonicated B. burgdorferi antigens of Korean isolates. Of 217 human patient's sera with unknown fever, the mean seropositivities in ELISA, done under the optimized conditions obtained in this study, were found to be about 8％ against B. burgdorferi sensu late, showing the highest seropositivity of 14.3％ against B. afzelii. In immunoblotting assay with ELISA-positive human sera, the major reactive bands were 41kDa (flagellin) which might be the indication of early infection, and 27kDa, 31kDa (OspA), 34kDa (OspB) which are the characteristics of late infection. Obtained results in this study might strongly indicate the possibility of B. burgdorferi infections in Korea.