• Parasites, Hosts and Diseases
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• 제56권1호
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• pp.61-70
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• 2018

We developed a Rapid Diagnostic Test (RDT) kit for detecting IgG/IgM antibodies against Zika virus (ZIKV) using monoclonal antibodies to the envelope (E) and non-structural protein 1 (NS1) of ZIKV. These proteins were produced using baculovirus expression vector with Sf9 cells. Monoclonal antibodies J2G7 to NS1 and J5E1 to E protein were selected and conjugated with colloidal gold to produce the Zika IgG/IgM RDT kit (Zika RDT). Comparisons with ELISA, plaque reduction neutralization test (PRNT), and PCR were done to investigate the analytical sensitivity of Zika RDT, which resulted in 100% identical results. Sensitivity and specificity of Zika RDT in a field test was determined using positive and negative samples from Brazil and Korea. The diagnostic accuracy of Zika RDT was fairly high; sensitivity and specificity for IgG was 99.0 and 99.3%, respectively, while for IgM it was 96.7 and 98.7%, respectively. Cross reaction with dengue virus was evaluated using anti-Dengue Mixed Titer Performance Panel (PVD201), in which the Zika RDT showed cross-reactions with DENV in 16.7% and 5.6% in IgG and IgM, respectively. Cross reactions were not observed with West Nile, yellow fever, and hepatitis C virus infected sera. Zika RDT kit is very simple to use, rapid to assay, and very sensitive, and highly specific. Therefore, it would serve as a choice of method for point-of-care diagnosis and large scale surveys of ZIKV infection under clinical or field conditions worldwide in endemic areas.

• Clinical and Experimental Pediatrics
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• 제50권12호
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• pp.1180-1187
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• 2007

Systemic lupus erythematosus (SLE) is an episodic, multi-system, autoimmune disease characterized by widespread inflammation of blood vessels and connective tissues and by the presence of antinuclear antibodies (ANAs), especially antibodies to native (double-stranded) DNA (dsDNA). Its clinical manifestations are extremely variable, and its natural history is unpredictable. Untreated, SLE is often progressive and has a significant fatality rate. The most widely used criteria for the classification of SLE are those of the American College of Rheumatology (ACR), which were revised in 1982 and modified in 1997. The presence of four criteria have been diagnosed as a SLE. Rashes are common at onset and during active disease. The oral mucosa is the site of ulceration with SLE. Arthralgia and arthritis affect most children and these symptoms are short in duration and can be migratory. Lupus nephritis may be more frequent and of greater severity in children than in adults. The initial manifestation of nephritis is microscopic hematuria, followed by proteinuria. The most common neuropsychiatric symptoms are depression, psychosis(hallucination and paranoia) and headache. CNS disease is a major cause of morbidity and mortality. Pericarditis is the most common cardiac manifestation. Libman-Sacks endocarditis is less common in children. The most frequently described pleuropulmonary manifestations are pleural effusions, pleuritis, pneunonitis and pulmonary hemorrhage. During the active phase ESR, CRP, gamma globulin, ferritin and anti-dsDNA are elevated. Antibodies to dsDNA occur in children with active nephritis. Antibodies to the extractable nuclear antigens (Sm, Ro/SS-A, La/SS-B) are strongly associated with SLE. Specific treatment should be individualized and based on the severity of the disease. Sepsis has replaced renal failure as the most common cause of death.

• 한국작물학회지
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• 제36권6호
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• pp.506-512
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• 1991

IAA에 대한 단크론 항체를 생산하고 이를 이용하여 생체중의 내생 IAA를 정량분석하기 위해 ELISA를 개발하고 본법을 사용하여 담배 종자 발아중 내생 IAA함량을 정량분석하였다. 그 결과는 1. IAA에 대한 단크론 항체 생산 세포주 3가지를 선발 작성하였으며 이 세포주로부터 생산되는 항체는 모두 IgG$_1$ 타입의 면역 글로블린이었다. 2. 상기 항체를 사용하여 ELISA를 수행하여 표준곡선을 작성하였던 바 검출 한계는 1pmol이었으며 검출 범위는 500pmol이었다 3. 표준곡선으로부터 작성한 Scatchard plot에 의한 친화 상수와 결합상수는 6.7$\times$$10^{-10}$ L/M과 6$\times$$10^{-10}$ L/M이었다. 4. 여러가지 IAA유사물질과 교차반응에 의해서 본 mAb는 특이성이 매우 높고 RIA에 의해서 고역가임을 확인하였다 5. 발아중인 담배종자로부터 면역측정에 의해서 내생 IAA를 정량분석하였다 6. 상기의 결과에 따라서 본 mAb를 이용하여 생체중의 내생 IAA를 간편하게 정밀 분석할 수 있음을 확인하였다.

• 한국가축번식학회지
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• 제14권2호
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• pp.93-100
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• 1990

These experiments were carried out to investigate the effect of rabbit anti-bovine cumulus cell antibodies on in vitro maturation of bovine follicular oocytes. Antisera to bovine cumulus cell were produced Japanese Ginat rabbit by repeated immunization of intact or solubilized bovine cumulus cell and purified by ammonium sulfate precipitation and Sepharose CL-4B protein-A affinity chromatography. The bovine cumulus cell-specific antibodies were confirmed by indirect ELISA. The results obtained in these experiments were summarized as follows : 1. The titer of the antibodies to cumulus cell determined by indirect ELISA using intact or solubilized bovine cumulus cell coated plates was very high in both intact and solubilized cumulus cells. Namely, the optical density at 1:12,800 dilution of antibodies was still significantly higher than that of non-immunized control serum. 2. When the follicular oocytes were treated with antibody to intact cumulus cells, the maturation rate of cumulus compacted and removed oocytes was ranged 47.6 to 59.1%. These value is significantly lower(p<0.05) than that(78.8%) of follicular oocytes cultured without the antibody. 3. the maturation rate of cumulus compacted and removed oocytes treated with antibody to solubilized cumulus cells was ranged 46.7 to 59.1%, significantly lower(p<0.05) than that(82.1%) of ooyctes cultured in antibody free medium. From above mentioned results, it could be said that cumulus cells promote nuclear maturation of follicular oocytes and that the beneficial effect of cumulus cells to the oocyte maturation is inhibited by the action of antibody to cumulus cells.

• Parasites, Hosts and Diseases
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• 제39권1호
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• pp.67-76
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• 2001

This experiment was focused on the characterization of anti- Toxoplasma monoclonal antibodies (mAbs) and the effect of mAbs on the parasite invasion of mouse peritoneal macrophages. Twenty eight mAbs including M110, M556, R7A6 and M62l were characterized by Ab titer, immunoglobulin isotyping and western blot pattern. Antibody titer (optical density) of 4 mAbs. Ml 10. M556. R7A6 and M62l. were 0.53,0.67, 0.45 and 0.39 (normal mouse serum; 0.19) with the same IgGl isotypes shown by Enzyme-linked immunosorbent assay (ELISA). Western blot analysis showed that Ml 10. M556. R7A6 and M62l reacted with the 33 kDa (p30),31 kDa (p28),43 kDa and 36 kDa protein. Immuno-gold labelling of mAbs M110, M556, R7A6 and M621 reacted with the surface membrane, dense granules and parasitophorous vacuolar membrane (PVM) , rhoptries and cytoplasm of tachyzoite, respectively. For in vitro assay, preincubation of tachyzoties with four mAbs, Ml 10, M556, R7A6 and M62l resulted in the decrease of the number of infected macrophages (p < 0.05) and the suppression of parasite multiplication at 18 h post-infection. Four monoclonal antibodies including Ml 10 (SAGI) were found to have an important role in the inhibition of macrophage invasion and T. gondii multiplication in vitro, and these mAbs may be suitable for vaccine candidates, diagnostic kit and for chemotherapy.

• Journal of Microbiology and Biotechnology
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• 제31권5호
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• pp.705-709
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• 2021

Porphyromonas gingivalis (P. gingivalis) is a major bacterial pathogen that causes periodontitis, a chronic inflammatory disease of tissues around the teeth. Periodontitis is known to be related to other diseases, such as oral cancer, Alzheimer's disease, and rheumatism. Thus, a precise and sensitive test to detect P. gingivalis is necessary for the early diagnosis of periodontitis. The objective of this study was to optimize a rapid visual detection system for P. gingivalis. First, we performed a visual membrane immunoassay using 3,3',5,5'-tetramethylbenzidine (TMB; blue) and coating and detection antibodies that could bind to the host laboratory strain, ATCC 33277. Antibodies against the P. gingivalis surface adhesion molecules RgpB (arginine proteinase) and Kgp (lysine proteinase) were determined to be the most specific coating and detection antibodies, respectively. Using these two selected antibodies, the streptavidin-horseradish peroxidase (HRP) reaction was performed using a nitrocellulose membrane and visualized with a detection range of 10³-10⁵ bacterial cells/ml following incubation for 15 min. These selected conditions were applied to test other oral bacteria, and the results showed that P. gingivalis could be detected without cross-reactivity to other bacteria, including Streptococcus mutans and Escherichia fergusonii. Furthermore, three clinical strains of P. gingivalis, KCOM 2880, KCOM 2803, and KCOM 3190, were also recognized using this optimized enzyme immunoassay (EIA) system. To conclude, we established optimized conditions for P. gingivalis detection with specificity, accuracy, and sensitivity. These results could be utilized to manufacture economical and rapid detection kits for P. gingivalis.

• BMB Reports
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• 제39권6호
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• pp.696-702
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• 2006

Recently three proteins, playing central roles in the bidirectional transport of urate in renal proximal tubules, were identified: two members of the organic anion transporter (OAT) family, OAT1 and OAT3, and a protein that designated renal urate-anion exchanger (URAT1). Antibodies against these transporters are very important for investigating their expressions and functions. With the cytokine gene as a molecular adjuvant, genetic immunization-based antibody production offers several advantages including high specificity and high recognition to the native protein compared with current methods. We fused high antigenicity fragments of the three transporters to the plasmids pBQAP-TT containing T-cell epitopes and flanking regions from tetanus toxin, respectively. Gene gun immunization with these recombinant plasmids and two other adjuvant plasmids, which express granulocyte/macrophage colony-stimulating factor and FMS-like tyrosine kinase 3 ligand, induced high level immunoglobulin G antibodies, respectively. The native corresponding proteins of URAT1, OAT1 and OAT3, in human kidney can be recognized by their specific antibodies, respectively, with Western blot analysis and immunohistochemistry. Besides, URAT1 expression in Xenopus oocytes can also be recognized by its corresponding antibody with immuno-fluorescence. The successful production of the antibodies has provided an important tool for the study of UA transporters.

• 생약학회지
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• 제41권4호
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• pp.275-281
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• 2010

To evaluate the immunomodulatory activity of a water extract (KMF-WE) of Korean mistletoe (Viscum album var. coloratum) fruit, we examined its ability to induce humoral and cellular immune response against keyhole limpet hemocyanine (KLH). Immunized mice with KLH admixed with KMF-WE (KLH/KMF-WE) showed significant induction of KLH-specific antibodies compared to mice immunized with KLH alone. The assay for determining isotypes of antibodies revealed that KMFWE augmented KLH-specific-IgG1 and -IgG2a production. In vitro T lymphocyte proliferation analysis against KLH revealed that the splenocytes of mice immunized with KLH/KMF-WE showed a significantly higher proliferative ability than those from mice immunized with KLH alone. The culture supernatants of splenocytes, which were harvested from mice immunized with KLH/KMF-WE, showed higher levels of both Th-1 type (IL-2, IFN-${\gamma}$) and Th-2 type (IL-4) cytokines in response to KLH stimulation compared to those from mice immunized with KLH alone. Also, in delayed-type hypersensitivity (DTH) assay, mice immunized with KLH/KMF-WE showed a significantly higher reaction to KLH than mice treated with KLH alone. These results suggest that KMF-WE possess immunoadjuvant activity to enhance both antigen-specific humoral and cellular immune responses against protein antigens (KLH).

• 대한본초학회지
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• 제25권4호
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• pp.23-29
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• 2010

Objectives : To investigate the immunological activities, we evaluated the combination ratio of Aconiti Lateralis Radix Preparata and Glycyrrhizae Radix (AG) on murine macrophage cell line (RAW 264.7) and ovalbumin/aluminium (OVA/Alum)-immunized mice. Methods : The cellular proliferation and the production of nitric oxide were examined in a macrophage cell line, RAW 264.7 cells, in the presence of the combination ratio of Aconiti Lateralis Radix Preparata and Glycyrrhizae Radix. C57BL/6 mice were immunized intraperitonially with ovalbumin/aluminium ($100{\mu}g/200{\mu}g$) on day 1, 8, and 15. The combination ratio of Aconiti Lateralis Radix Preparata and Glycyrrhizae Radix (1 g/kg/day) was orally administrated for 3 weeks. On day 22, splenocyte and plasma were collected for mitogen-induced proliferation, lymphocyte subpopulation by flow cytometry and measurement of AST (Aspirate aminotransferase), ALT (Alanine aminotransferase), and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes). Results : Aconiti Lateralis Radix Preparata treatment had no influence on immune responses. The proliferation and NO production of macrophage and proliferation of splenocyte were increased as the higher ratio of Glycrrhizae Radix. The proliferation of splenocyte, lymphocyte subpopulation and production of antibody (total IgM, OVA-specific IgG and OVA-specific IgG1) were increased as the higher ratio of Glycrrhizae Radix on OVA-immunzed mice. Conclusions : These results suggest that the higher ratio of Glycyrrhizae Radix can increase immunological activities such as NO production in RAW264.7 cells, splenocyte proliferation and immunoglobulin production in OVA-immunized mice.

• 대한수의학회지
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• 제43권1호
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• pp.129-137
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• 2003

Porcine reproductive and respiratory syndrome (PRRS) is characterized by reproductive failures in sows and respiratory problems in piglets. The nucleocapsid(N) protein, encoded by the open reading frame 7 (ORF7) gene, is known to be the most abundant and antigenic protein in PRRS virus. Therefore, it was suggested that the N protein could be a suitable candidate for the detection of PRRS virus-specific antibodies and diagnosis of PRRS. In the present study, the ORF7 gene encoding the N protein was cloned and expressed as a fusion protein with the glutathione S-transferase (GST) in Escherichia coli. The resulting GST-N recombinant protein was used as an antigen for an indirect sandwich enzyme-linked immunosorbent assay (i-ELISA). Expressed GST-N recombinant protein was migrated at 41 kDa and reacted with ORF7-specific monoclonal antibody by Western blotting. In order to increase the specificity of the ELISA for the detection of PRRS virus-specific antibodes, an i-ELISA was developed using an anti-GST antibody as a capture antibody. The sensitivity and specificity of developed i-ELISA were 92% and 96%, respectively. Based on these results, it was suggested that the i-ELISA is a simple and rapid test for screening a large number of swine sera for the anti-PRRS virus antibodies.