• Tuberculosis and Respiratory Diseases
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• v.59 no.1
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• pp.47-52
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• 2005

Background : Chlamydia pneumoniae is a clinically important pathogen, the diagnosis of such infection being based mainly on serology. Microimmunofluorescence (MIF) is the current standard diagnostic method, but is subjective and time-consuming, so the authors tested the serology of chronic cough patients using an EILSA method for the Chlamydial antibody, which is a more objective method, and compared the results with those of the standard method. Method : Thirty-five patients, who visited Kangwon National University Hospital between August 2003 and July 2004, were evaluated. A MIF and ELISA tests were used to determine C. pneumoniae antibody titers. Nasopharyngeal aspirates were examined by polymerase chain reaction (PCR). The Spearman rank correlation test was used for data analysis. Results : Sensitivities of ELISA for IgG, IgA and IgM, as judged by MIF, were 84.0, 84.0 and 40.0% and the specificities were 60.0, 60.0 and 96.7%, respectively. Three patients were Chlamydia PCR positive. Conclusion : ELISA can be a useful tool for studying the seroprevalence of Chlamydia pneumoniae. However, further studies will be required prior to its clinical use.

• Journal of Microbiology and Biotechnology
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• v.18 no.9
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• pp.1606-1612
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• 2008

Interleukin (IL)-32 is a recently identified proinflammatory cytokine that is one of the IL-18 inducible genes, and plays an important role in autoimmune and inflammatory diseases. We produced antibodies against IL-32 and studied the expression of IL-32 in human stomach cancer. We detected IL-32 secreted from K-562 cells which were stably transfected with IL-32 and in the sera of stomach cancer patients by a sandwich ELISA using a monoclonal antibody KU32-52 and a polyclonal antibody. In order to optimize a sandwich immunoassay, recombinant IL-32a was added, followed by the addition of a biotinylated KU32-52 into microtiter plate wells precoated with a goat anti-IL-32 antibody. The bound biotinylated KU32-52 was probed with a streptavidin conjugated to HRP. This sandwich ELISA was highly specific and had a minimal detection limit of 80 pg/ml (mean${\pm}$SD of zero calibrator) and measuring up to 3,000 pg/ml. This ELISA showed no cross-reaction with other cytokines such as hIL-1$\alpha$, hIL-1$\beta$, hIL-2, hIL-6, hIL-8, hIL-10, hIL-18, and hTNF-$\alpha$. Intra-assay coefficients of variation were 18.5% to 4.6% (n=10), and inter-assay coefficients were 23% to 9% (n=10). The average IL-32 level in the sera of 16 stomach cancer patients (189 pg/ml) was higher than that of 12 healthy control men (109 pg/ml). Our results indicate that serum IL-32 level can be detected by using an established ELISA, and that this immunoassay and mAb KU32-09 specific for immunohistochemistry can be used in the detection of expressed and secreted IL-32 in stomach cancer patients.

• Korean Journal Plant Pathology
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• v.14 no.2
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• pp.130-135
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• 1998

This study was carried out to detect cymbidium mosaic potexvirus (CymMV) and odontoglossum ringspot tobamovirus (ORSV) in cultivated orchid plants in Korea. The standard double antibody sandwich enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR) were carried out for detection of the viruses in the collected orchid samples. ELISA was suitable for massive-scale diagnostic method for virus detection in orchids. RT-PCR was rapid, time-saving and reliable detective method, and detection limit data showed that RT-PCR was 103 times more sensitive than ELISA. Of the 321 individual orchids representing 5 orchids genera tested by the ELISA, CymMV and ORSV were detected in 15.6% and 22.4%, and mixed infection of the both viruses with 4.9%, respectively. Of the Cymbidium plants tested, cultivated plants showed 52.5% virus infection rate with either CymMV or ORSV and both viruses.

• Food Science of Animal Resources
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• v.30 no.1
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• pp.36-41
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• 2010

The Hen's egg is widely used in many processed foods as an ingredient and is one of the most prevalent food allergens in children. To detect egg proteins in processed foods, we developed a competitive indirect enzyme-linked immunosorbent assay (ciELISA) using an anti-ovomucoid (OM) antibody, which was produced by immunization of rabbits with OM, the most heat-stable component of the egg proteins. The detection limit of this quantitative assay system was 30 ng/mL. Cross-reactivity of the anti-OM antibody toward OM, ovalbumin, skim milk, casein, whey protein isolate, and isolated soy protein was 100, 0.4, 0.2, 0.04, 0, and 0%, respectively. In the spike test of egg white powder in milk replacer, commercial sausage, and in-house sausage, the assay recoveries ($mean{\pm}SD$) were $129{\pm}13.7%$, $73.9{\pm}12.5%$, and $65.5{\pm}13.6%$, respectively. When egg white in a commercial crab meat analog and sausage was determined by ciELISA, the assay recovery was found to be 108% and 127%, respectively. The combined results of this study indicate that this novel ciELISA for OM detection could be applied for the quantification of hen's egg proteins in processed foods.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.43 no.4
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• pp.228-233
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• 1998

Seed storage proteins have been used for studying biochemical genetics and end-use quality aspects. We conducted enzyme-linked immunosorbent assay (ELISA) and one-dimensional SDS-PAGE (1D SDS-PAGE) to evaluate different cereal crop species and Korean wheat lines for rye secalin proteins. The antisecalin antibody showed consistent specificity for rye secalin with little cross-reactivity to gliadins. Immunological cross-reactivities measured by the ELISA technique using competition assay showed significant differences of absorbance among rye, triticale, wheat-rye translocated wheat and non-translocated wheat. The absorbance values were lowest in rye followed by triticale, translocated wheat and non-translocated wheat. The ELISA for discrimination of wheat-rye translocation on the basis of antigen-antibody reactivity showed that none of the Korean wheat lines possessed 1RS and secalin proteins. The competitive ELISA experiment demonstrated specific determination for secalin that was originated from rye chromosomal parts. The result of 1D SDS-PAGE for identifying rye secalin subunits showed all three rye specific secalin protein subunits (75 KDa, 45 KDa, and 40 KDa) for rye and triticale, and 1RS specific secalins (45 KDa and 40 KDa) for 1AL/1RS and 1BL/1RS translocated wheats. All Korean wheats were lacking 1RS of rye chromosome and secalin.

• Korean Journal of Veterinary Service
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• v.32 no.4
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• pp.293-298
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• 2009

Porcine respiratory coronavirus (PRCV) is antigenically related to transmissible gastroenteritis virus (TGEV). Differential serological diagnosis between PRCV and TGEV infection is not possible with the classical sero-neutralization test. Infection with PRCV or TGEV induces antibodies which neutralize both viruses to the same titer. However, the enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) can differentiate between PRCV and TGEV infection. This study was carried out to investigate the prevalence of PRCV infection of swine in Gyeongnam province. A total of 391 serum samples from 37 herds in Gyeongnam were examined for antibody to PRCV using blocking ELISA. All serum samples were collected from 130- to 150-day-old pigs between August and December 2006. By ELISA, 182 out of 391 sera tested (46.5%) and 29 out of 37 sample herds (78.4%) were positive against PRCV. Our data suggested that seropositive herds for PRCV are distributed diffusely throughout Gyeongnam. The PCR methods were established to diagnose PRCV spike protein (S) gene. PCR were conducted to identify the PRCV genome against 150 pigs in PRCV antibody positive herds.

• Parasites, Hosts and Diseases
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• v.50 no.2
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• pp.99-102
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• 2012

Serum samples, 100 in the total number, were collected from different laboratories in Tehran, Iran and tested for anti-Toxoplasma specific IgG and IgM antibodies using indirect immunofluorescent antibody test (IFAT). Using the IgG (chronic) and IgM (acute) positive samples, the IgG avidity test was performed by ELISA in duplicate rows of 96-well microtiter plates. One row was washed with 6 M urea and the other with PBS (pH 7.2), then the avidity index (AI) was calculated. Sixteen out of 18 (88.9%) sera with acute toxoplasmosis showed low avidity levels ($AI{\leq}50$), and 76 out of 82 (92.7%) sera in chronic phase of infection showed high avidity index (AI>60). Six sera had borderline ranges of AI. The results showed that the IgG avidity test by ELISA could distinguish the acute and chronic stages of toxoplasmosis in humans.

• Parasites, Hosts and Diseases
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• v.21 no.2
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• pp.246-256
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• 1983

As epidemiological parameters of human paragonimiasis, the positive rates of intradermal test and the sputum/stool ekaminations have long been employed in population surveys. However, both the specificity of the intradermal test and the sensitivity of sputumjstool examination have been gradually declined as the endemicity was lowered; thus the gap between above two parameters widened. In such context, the development of a new epidemiologic parameter or tool which makes it possible to accurately discriminate the active paragonimiasis cases was necessary. In the present study, the detection rate of Paragonimus-speclac IgG antibody by micro-ELISA was evaluated as an indicator of epidemiologic status of human paragonimiasis in a population. A total of 4, 285 students and inhabitants living in Bukpyeong Myeon and Bukil Myeon, Haenam Gun, Jeonlanam Do was surveyed in October, 1983 by intradermal test first. Out of them, 244 cases (5.7%) were found positively reacted to VBS antigen of F. westermani. Out of 168 positive reactors, 7 cases (4.2%) were egg positive either by two times of sputum examination or by one stool examination. That indicated that only 0. 16% of total surveyed were confirmed as active paragonimiasis by egg detection. When sera collected from 239 positive reactors of Intradermal test were tested by micro-ELISA for their specific IgG antibody, 40 cases(16.7%) were found to be positive. All of 7 egg positive cases were again positive for specific IgG antibody. Among remaining 232 intradermal test positive cases, 33 cases were positive for IgG antibody. In contrast to those, none of 42 positive reactors to intradermal test for Cloncrchis and of 128 intradermal test negative cases showed positive for Paragcnimus-specIfic IgG antibody. The rate of specific IgG antibody as detected by micro-ELISA appeared to be increased with the wheal size of the intradermal test. When the wheal sixte was over 13mm in diameter, about 50% of them were positive for specific IgG antibody. Thirty-one specific antibody positive cases were clinically evaluated by laboratory examinations (repeated sputum examination, peripheral eosinophil count and chest roentgenogram) and by history taking. Out of them 24 cases were associated with one or more positive laboratory findings: thus considered as active paragonimiasis cases. Out of 7 lab. finding-free cases 3 revealed past history of typical paragonimiasis symptoms, suggesting that they were in chronic or in convalescent stages. The remaining 4 cases were considered as either mild or ectopic infection cases; the possibility of cross-reaction with other helminthiases could not be ruled out. From the above results, it was inferred that the detection of Paragonimus-specIfic IgG antibody by micro-ELISA was very much helpful in detecting the active cases as well as in proper evaluation of the endemicity of human paragonimiasis in a population. The convenience of mass haildling of sera in micro-ELISA was considered another superiority as an epidemiologic tool.

• Parasites, Hosts and Diseases
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• v.34 no.4
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• pp.233-238
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• 1996

The seroepidemiologic studies on antral-ToxopIasma antibody titers were carried out using ELISA and indirect latex agglutination test. Among 899 sera prepared from pregnant women, 39 cases (4.3%) revealed positive reaction and 218 sera from middle school students showed 4 positive reaction (1.8%) by ELISA. By LAT (newly established by National Veterinary Research Institute, Korea), the sera of 7 pregnant women (0.8%) showed positive reaction. When 80 sera showing ≥ 1 :8 by LAT were used for comparing the results obtained from LAT and Toxotest-MT (Eikon Chemical Co., Japans, 7 cases and 8 sera were positive, respectively. All of 11 sera of proven toxoplasmosis patients showed positive reaction in both tests. Overall proportion of agreement between LAT kit and Toxotest-MT was 0.94 (K-index : 0.632, p < 0.01), and LAT was considered to be useful for the screening of toxoplasmosis.

• Research in Plant Disease
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• v.18 no.3
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• pp.231-235
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• 2012

Indian citrus ring spot virus (ICRSV) is known to cause a serious disease to citrus, especially to Kinnow mandarin, the popular cultivated citrus species in India. In this study, we developed diagnostic systems based on enzyme-linked immunosorbent assay (ELISA). In order to generate antibodies against ICRSV coat protein, we overexpressed the coat protein in Escherichia coli using the pET15b expression vector containing an optimized ICRSV coat protein gene. The recombinant ICRSV coat protein was overexpressed as soluble form at $37^{\circ}C$ upon IPTG induction. The protein was purified to 95% in purity by Ni-NTA column chromatography. The purified protein was immunized to rabbit for the generation of polyclonal antibody (PAb). The PAb showed a specific immunoreaction to recombinant ICRSV coat protein in western blot analysis and ELISA. Diluted rabbit antisera (10,000 fold) could detect less than 10 ng and 5 ng of the target protein in western blot and ELISA analysis, respectively.