• Archives of Pharmacal Research
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• 제19권6호
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• pp.486-489
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• 1996

A simple, rapid, specific and sensitive method for the detection of serum hepatitis C virus (HCV) RNA using the reverse transcription-polymerase chain reaction (RT-PCR) technique without conventional RNA extraction was developed. HCV template RNA from serum was obtained by boiling the serum at $95^{\circ}C$ for 2 min, cooling rapidly in ice and removing the proteins by cetrifugation. RT-PCR amplifications including the reverse transcription and first PCR amplification were performed in one vessel containing both of reverse transcriptase and Taq DNA polymerase. The detection of HCV RNA from $10^{-3}{\mu}l$. serum was possible with this method. The suitability of this method for clinical analysis was evaluated by assaying HCV RNA in 225 patient samples including anti-HCV antibody negatives (13 samples) and positives (212 samples) by enzyme-linked immunosorbent assay test (ELISA). Detections of HCV RNA with this method were in 4 of 13 anti-HCV antibody negative samples (30.8%) and 95 of 212 positive samples (44.8%). The present method can be completed in 1 hr and has a wide range of application for the clinical utilities to determine the viral RNAS.

• International Journal of Industrial Entomology and Biomaterials
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• 제9권2호
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• pp.265-267
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• 2004

The silkworm, Bombyx mori L. is prone to infection of various pathogenic organisms. Pebrine, one of the deadliest disease of silkworm caused by highly virulent parasitic microsporidian, Nosema bombycis has been understood since long. Infections of the disease range from chronic to highly virulent and can result in complete lose to the sericulture industry. Several strains and species of microsporidians have since been isolated from the infected silkworms. A new microsporidian spore was isolated from Lamerin breed of the silkworm B. mori have been studied under scanning electron microscope, found to be different in spore size (length 4.36$\pm$0.06 ${\mu}{\textrm}{m}$, width 2.14$\pm$0.01${\mu}{\textrm}{m}$) and shape (ova cylindrical with slight depression) from standard strain N. bombycis (length 3.08$\pm$0.21 ${\mu}{\textrm}{m}$, width 2.01$\pm$0.05 ${\mu}{\textrm}{m}$ and ovidal respectively). In immunological test, the silkworm breed Lamerin isolated micrisporidian spore does not react to different antibody (N. bombycis, M$_{11}$ and M$_{12}$) sensitized latex particle and thus appeared to be a different strain of microsporidian parasitic to the Lamerin breed of the silkworm B. mori.i.i.

• Journal of Preventive Medicine and Public Health
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• 제34권4호
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• pp.444-446
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• 2001

Objectives : Toxoplasmosis is a member of the zoonosis group and may cause congenital infection . Antibody positive rates of toxoplasmosis were examined in high school students in Cheju, Korea to facilitate the study aim of examining the seroprevalence of Toxoplasma gondii in school teachers of child-bearing age in Cheju Island. Methods : The study population comprised teachers of child-bearing age in primary, middle and high schools, aged 35 years and younger, who wished to be tested for Toxoplasma gondii antibodies (IgG) by the indirect latex agglutination test (ILA) and the enzyme-linked immunosorbent assay (ELISA) method. Results : The overall antibody positive rate was 3.8% in the study subjects (n=314), a rate which showed no significant difference due to birth place, history of bringing up pets, or history of contacting a cat. Conclusion : We confirmed that the seroprevalence of Toxoplasma gondii in a population of child-bearing teachers in Cheju Island was the same as that previously reported in other parts of Korea.

• Bulletin of the Korean Chemical Society
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• 제22권9호
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• pp.953-957
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• 2001

The specificity of conjugation site for coating ligands was investigated using toluene-bovine serum albumin (BSA) conjugates in which BSA was conjugated at the position of o-, m-, and ${\rho}-toluic$ acid. Toluene-BSA conjugated at ${\rho}-position$ showed a binding activity of about 89-95%, whereas, those conjugated at o- and m-position of toluene exhibited a binding activity of 5 and 11%, respectively. On the basis of the above result, coating ligands with various proteins (OVA, BSA, KLH) were compared by conjugating with $\rho-toluic$ acid, and toluene-KLH was considered as the best coating ligand for this ELISA. Indirect competitive ELISA method was developed using anti-toluene antibody and $\rho-position$ conjugated toluene-KLH. The dose-response curve constructed after kinetic and optimization studies showed a 1${\times}$10-4 - 1${\times}$102 mM detectable response range with 0.1 ${\mu}M$ detectability. In specificity test of the antibody, the binding capabilities of aromatic compounds substituted with nitro-, alkyl-, chloro-, and hydroxyl group were larger rather than those of aromatic compounds (benzene, toluene and xylene) themselves. Also, tests with soil and water samples that had been spiked with toluene resulted in 102.7-113.7% recovery.

• 한국어병학회지
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• 제36권2호
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• pp.389-394
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• 2023

Infectious hematopoietic necrosis virus (IHNV) is s significant viral pathogen affecting cultured rainbow trout (Oncorhynchus mykiss) in Korea. In this study, five monoclonal antibodies (mAbs) (IHNV-1, 2, 3, 4, and 5) were produced using purified IHNV. Reactivities of these mAbs were analyzed by western blot (WB), enzyme-linked immunosorbent assay (ELISA), and indirect fluorescent antibody test (IFAT). These mAbs recognized glycoprotein (69 kDa, IHNV-1), nucleocapsid protein (39 kDa, IHNV-3, 4, and 5), or phosphoprotein (27 kDa, IHNV-2) of IHNV by WB analysis. ELISA results indicated that these five mAbs were specific to IHNV without showing any cross-reactivity against other fish viruses (hirame rhabdovirus, infectious pancreatic necrosis virus, and viral hemorrhagic septicemia virus). IFAT demonstrated specific fluorescence signals of IHNV-infected epithelioma papulosum cyprini (EPC) cells, whereas no reactivity of normal EPC cells was observed. These mAbs can be very useful for immuno-diagnosis of IHNV infection.

• 한국동물위생학회지
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• 제22권3호
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• pp.221-237
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• 1999

To investigate the specificity of rapid slide agglutination test for pullorum-gallinarum diseases and to obtain a basic data for avian salmonellosis control, salmonella isolation was peformed for the layer chickens positively reacted in pullonlm-typhoid agglutination test. The biochemical, serological and antimicrobial properties of the isolates were examined. The results obtained through this study were summarized as follows; 1. Of 2,384 chickens tested by the agglutination test, 606 chickens (25.4%) were positive reactors. 154 of 606 reactors and 49 of the non-reacting chickens were investigated for salmonella isolation, resulting in isolation of 68 strains of salmonellae from 27 chickens. 2. By organs, the isolation frequency from liver, cecum, spleen, ovary and gall bladder showed 8.9% (18 strains), 8.9% (18 strains), 7.4% (15 strains), 4.4% (9 strains) and 3.9% (8 strains), respectively. 3. By culture medium the combination of selenite broth and MacConkey agar revealed the highest isolation rate and the enrichment culture by delayed secondary enrichment culture method was found the most effective for salmonella isolation. 4. The serotypes of 68 salmonella isolates were identified as 3 strains of S pullorum, 24 strains of S gallinarum, 15 strains of S typhimurium, 8 strains of S enteritidis, 7 strains of S paratyphi A, 5 strains of S typhimurium and 6 strains of the other salmonellae. 5. The serotypes of 8 salmonella strains isolated from 49 chickens non-reacting in pullorum-typhoid agglutination test were identified as 3 strains of S typhimurium and 5 strains of S infantis. 6. When 24 chickens of which 68 strains of salmonellae isolated were examined by microplate agglutination test, the average antibody titer for pullorum antigen was $2^{5.25}$. The chickens at antibody titer between $2^3$ and $2^5$ showed the higher frequency of isolation as compared with the chickens at the other titers. 7. When salmonella isolates were tested the antimicrobial drug sensitivity by disk diffusion method, S paratyphi A were highly sensitive by 100% to ATM and GM, S typhimurium, by 88% to AM, CIP, IMP and TN, S infantis, by 100% to AM, CRO, ENR and PIP, S enteritidis,by 100% to IMP and PIP, S pullorum, by 100% to ATM, CRO, ENR and PIP and S gallinarum, by 92% to CRO, CIP and PIP.

• Parasites, Hosts and Diseases
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• 제31권2호
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• pp.141-148
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• 1993

폐흡충증의 진단을 위하여 근자에 효소면역측정 법(ELISA)이 널리 쓰이고 있다. 이 논문에서는 폐흡충 항원에 대한 단클론항체를 제조하고 이를 효소면역억제측정 법 (ELISA-Inhlbltlon test)에 응용하여 효소면역측정 법의 특이도를 더욱 높이고자 하였다. 폐흠충 성충의 수용성 항원으로 면역한 BALB/c 마우스의 비장세포와 형질세포종세포를 융합하여 림프잡종세포를 만들고 이 중에서 폐흡충 항원에 대한 특이 단클론항체를 생성하는 하나의 림프잡종세포를 선택하였다. 이는 Pwa-14라고 이름지워졌는데, EITB상 28 kDa, 42.5 kDa, 89 kDa, 120.5 kDa 항원대에 반응하였으며, 간접 면역항체법으로 반응하는 항원의 위치가 난황선임을 확인하였다. 폐흡충 항원을 사용한 통상적인 효소면역측정법에 폐흡충 감염자 22명의 혈청은 모두 양성 반응을 보였으며, 간흡충 감염자 40명 중 5명(12.5%), 유구낭충 감염자 26명 중 3명(7.7%)은 양성을 나타내 교차 반응을 보였다. 이 때, 스파르가눔 감염자 10명 및 정상 대조군에서는 양성 반응을 보이는 혈청이 없었다. 특이 단세포군항체를 이용한 효소면역 억제측정법으로 측정한 결과, 폐흡충 감염자는 모두 양성반응을 보였으나, 유구낭충, 스파르가눔 감염자의 혈청은 교차 반응을 보이지 않았다. 이상의 결과로 보아 폐흡 충의 면역 진단 시, 폐흠충 특이 단클론항체를 이용한 효소면역 억제측정 법은 통상적인 효소면역측 정법에 비하여 더 높은 특이도를 보임을 알 수 있었다.

• 한방재활의학과학회지
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• 제19권1호
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• pp.39-55
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• 2009

Objectives : This study was designed to evaluate effects of electroacupuncture and Cervi Pantortichum Cornu pharmacopuncture on pain and nerve regeneration after crush injury of sciatic nerve in rats. Methods : We divided the subjects into 5 groups : control group, acupuncture treated group, electroacupuncture treated group, Cervi Pantortichum Cornu pharmacopuncture treated group, electroacupuncture and Cervi Pantortichum Cornu pharmacopuncture group. Hot plate test and degree of Substance P antibody in each group were observed at the 1st, 2nd, 3rd and 5th day. Sciatic function index and change of BDNF immunoreactivity had been estimated at the 7th, 14th, 21st day respectively. Atrophy of the gastrocnemius at the 21st day was observed. Results : All the experimental groups showed significant changes in hot plate test, degree of Substance P antibody, sciatic function index and change of BDNF immunoreactivity compared with control group. Especially, when electroacupuncture and Cervi Pantortichum Cornu pharmacopuncture used together, results were more effective than other groups. Recovery of the gastrocnemius was also the same. Conclusions : It is suggested that electroacupuncture and Cervi Pantortichum Cornu pharmacopuncture may play a significant role on pain decrease and nerve regeneration after crush injury of sciatic nerve in rats and these two treatments could be more effective when used together.

• Parasites, Hosts and Diseases
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• 제52권2호
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• pp.143-149
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• 2014

To evaluate the seroprevalence against circumsporozoite protein (CSP) of Plasmodium vivax in sera of Korean patients, the central repeating domain (CRD) of CSP was cloned and analyzed. From the genomic DNA of patient's blood, 2 kinds of CSPs were identified to belong to a VK210 type, which is the dominant repeating of GDRA(D/A)GQPA, and named as PvCSPA and PvCSPB. Recombinantly expressed his-tagged PvCSPA or PvCSPB in Escherichia coli reacted well against sera of patients in western blot, with the detecting rate of 47.9% (58/121), which included 15 cases positive for PvCSPA, 6 cases positive for PvCSPB, and 37 cases for both. The mixture of PvCSPA and PvCSPB was loaded to a rapid diagnostic test kit (RDT) and applied with the same set of patient sera, which resulted in detection rates of 57.0% (69/121). When the protein sequences of PvCSPA were compared with those of P. vivax in endemic regions of India and Uganda, they were compatibly homologous to PvCSPA with minor mutations. These results suggested that the recombinant PvCSPA and PvCSPB loaded RDT may be a milestone in latent diagnosis which has been a hot issue of domestic malaria and important for radical therapy in overlapped infections with P. falciparum in tropical and subtropical areas. During the biological process of malarial infection, exposure of CSP to antigen-antibody reaction up to 57.0% is the first report in Korea.

• 한국축산식품학회지
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• 제24권2호
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• pp.182-188
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• 2004

Lactoperoxidase(LPO)를 농도를 측정하기 위한 ELISA을 개발하기 위해 LPO로 면역시킨 갈색 산란계의 계란에서 형성된 anti-LPO IgY 항체를 분리 정제하고, 분리된 anti-LPO IgY 항체의 특이성을 ELISA 와 double immunodiffusion 방법으로 조사한 후 indirect ELISA 방법을 이용한 표준곡선을 만들었다. 분리 정제된 anti-LPO IgY항체의 titer는 1:520,000이며, ELISA와 double immunodiffusion 방법 모두에서 $\alpha$-lactalbumin, $\beta$-lactoglobulin, casein 및 lysozyme하고는 교차반응을 하지 않고 LPO만 높은 특이성을 갖는 항체로 나타났다. Indirect ELISA방법에서 LPO의 coating 농도는 0.25 $\mu\textrm{g}$/mL이며 anti-LPO IgY 최적 희석배수는 1:8,000으로 나타났다. Indirect ELISA 방법으로 LPO를 측정할 수 있는 표준곡선에서 민감도의 범위는 0.0l-l $\mu\textrm{g}$/mL로 나타났다.