• IMMUNE NETWORK
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• 제1권1호
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• pp.1-6
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• 2001

The identification of tumor-specific antigens has represented a critical milestone in cancer diagnosis and therapy. Clinical research in this area for leukemia has also been driven over the past few decades by the hope that surface antigens with restricted tissue expression would be identified. Disappointingly, only a small number of the leukemic antigens identified to date, meet sufficient criteria to be considered viable immunophenotypic markers. In this paper, we nominate anti-JL1 monoclonal antibody as an immunodiagnostic and immunotherapeutic candidate for leukemia. The JL1 molecule appears to be a novel cell surface antigen, which is strictly confined to a subpopulation of limited stages during the hematopoietic differentiation process. Despite the restricted distribution of the JL1 antigen in normal tissues and cells, anti-JL1 monoclonal antibody specifically recognizes various types of leukemia, irrespective of immunophenotypes. On the basis of these findings, we propose JL1 antigen as a tumor-specific marker, which shows promise as a candidate molecule for diagnosis and immunotherapy in leukemia, and one that spares normal bone marrow stem cells.

• 대한바이러스학회지
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• 제26권1호
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• pp.77-90
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• 1996

Nucleocapsid protein (NP)which exists in the particle of hantavirus and surrounds the viral RNA genome is one of the major structural proteins and plays role of antigen to elicit the antibody detected predorminantly right after infection of the virus in the patients of hemorragic fever with renal syndrome (HFRS)or experimental animals. NP is important target antigen in serological diagnostic system of HFRS utilizing whole antigens from the native virus particle, such as IFA, ELISA and Western blotting. Therefore, the preparation of this protein in the level of higher quantity and purity is desirasble for developed dianosis of the disease. The purpose of this study is the cloning of NP gene which exists in the S genome segment of Maaji (MAA) virus and expression of the gene to obtain qualified, genetically engineered NP to be utilized as an immunodiagnostic antigen. First of all, for the purpose of amplifing the MAA-NP gene by PCR, the specific primers were built from the known nucleotide sequence of Hantaan viral NP gene. The viral cDNA of the NP gene was synthesized by using the primers and RNase $H^-$ AMV reverse transcriptase. Thereafter, using this cDNA as a template, the NP gene was amplified specifically by Taq DNA polymrerase. The pT7blue (R)T-overhang vector systems were used for cloning of the amplified NP gene. The expression system was consisted of BL21 (DE3)pLysS and pET16b as a host and a plasmid repectively. Into Ndel site of pET16b, NP gene was ligated with cohesive end for the expression. Insertion of NP gene in the plasmid was confirmed by PCR and mini prep methods. For expression, IPTG was used and the expressed protein was characterized by Western blotting. The MAA-NP was expressed as the form of inclusion body (insoluble fraction)and the protein purified by affinity and metal chealating columns reacted specifically with the sera from patients of HFRS as to be tested by ELISA and Western blotting.

• 한국동물학회지
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• 제38권3호
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• pp.382-387
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• 1995

항-AFP 단일 클론 항체를 생산한 다음, 이를 이용한 noncompetitive ELISA 방법으로 정상인과 간암 및 그 밖의 간질환 환자의 혈청내 AFP농도를 측정해 본 결과 간암진단 방법으로는 혈청 AFP농도 측정이 필수적임이 확인되었다. 또한 간암 및 그 밖의 간질환 환자의 조직에 대한 항-AFP-항체의 반응성을 immunperoxidase 방법과 indirect immunofluorescence 방법으로 검정해 본 결과, 간암조직세포 및 일부 간질환 조직세포에서 항-AFP-항체에 대해 양성반응을 나타내었다. 그러나 그 반응성의 정도는 간암조직세포에서 보다 간암조직 주위의 비신생 간세퐁서 더욱 높았다. 그러므로 간암 진단에 있어서 AFP항원을 면역조직화학적으로 검정하는 방법은 적합하지 않았다.

• Parasites, Hosts and Diseases
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• 제47권2호
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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.

• Parasites, Hosts and Diseases
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• 제25권1호
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• pp.7-12
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• 1987

The indirect fluorescent antibody(IFA) test was used to detect serum IgG and IgM antibodies to Trichomonas vaginalis in 31 vaginal trichomoniasis, 7 candidiasis and in 20 non-infected healthy women with antigen prepared from axonic culture of Trichomenas vaginalis isolated from vulvovaginitis patient. The results were as follows: 1. In 31 vaginal trichomoniasis the positive reactions of IgG antibody were 27 in the 1/8 dilution or higher and :l in the 1/4 dilution whereas in healthy women the reaction showed significantly low as in the 1/4 dilution or below. 2. The sensitivity and specificity of IFA test for IgG antibody to trichomonad antigen in this study were 87.1% and 100%, respectively. 3. No significant difference of IgM antibody levels between vaginal trichomoniasis and healthy women was observed. 4. No relation between the levels of IgG and IsM antibodies to trichomonad antigen by IFA test was observed. 5. No relation between the time lapse and the level of serum IgG antibodies in IFA test of vaginal trichomoniasis was regarded. In conclusion the present study suggests that IFA test in trichomoniasis could be a useful tool for detection of anti-trichomonad IgG antibodies and applicable as an immunodiagnostic method.

• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1180-1184
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• 2006

The capsule that surrounds Yersinia pestis cells is composed of a protein-polysacchride complex; the purified protein component is fraction I (F1) antigen. We report the cloning of the cafl gene and its expression in Escherichia coli using the vector pETl02/D-TOPO and the F1-specific monoclonal antibody. The recombinant F1 (rF1) antigen had a molecular size of 17.5 kDa, which was identical to that of the F1 antigen produced by Y. pestis. Recombinant F1 protein was found to react to polyclonal antiserum to Y. pestis Fl. Recombinant F1 was purified by ProBond purification system and induced a protective immune response in BALB/c mice challenged with up to 10$^5$ virulent Y. pestis. Purified rF1 protein was used in an ELISA to evaluate the ability of a method to detect antibodies to Y. pestis in animal sera. These results strongly indicated that the rF1 protein is a suitable species-specific immunodiagnostic antigen and vaccine candidate.

• Parasites, Hosts and Diseases
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• 제43권1호
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• pp.15-18
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• 2005

The diagnosis of human hydatidosis is primarily made using radiological and serological methods. Radiological methods are generally of low specificity and serological methods lack sensitivity, especially for pulmonary disease. In this study the capabilities of a new rapid test, the hydatid antigen dot immunobinding assay (HA-DIA), which was developed for the diagnosis of pulmonary hydatidosis, were studied and compared with another immunodiagnostic method, indirect hemagglutination (IHA). The study subjects included 18 patients, 9 women, 9 men; range 7 to 63 years; mean 30 years, with surgically proven pulmonary hydatidosis, a control group comprised of 14 patients; viral respiratory infections (1), cirrhosis (2), connective tissue disease (2), taeniasis (3), and 6 healthy donors. We found that the HA-DIA test had a sensitivity of $67\%$ and specificity of $100\%$, and that the IHA test had a sensitivity of $50\%$ and specificity of $100\%$. We conclude that HA-DIA is a simple, rapid, low cost assay that does not require instrumentation and has a higher sensitivity than IHA for the diagnosis of pulmonary hydatidosis.

• Tuberculosis and Respiratory Diseases
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• 제39권1호
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• pp.1-6
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• 1992

The diagnosis of tuberculosis is usually established using staining and culturing techniques. Fluorescent stains have improved the sensitivity of direct microscopy. Improved culture media coupled with radiometric means of detecting early mycobacterial growth have shortened the time needed for cultural diagnosis. Rapid immunodiagnostic techniques based on the detection of mycobacterial antigen or of antibodies to theses antigens have not, however, come into widespread clinical use. The DNA or RNA hybridization tests with labeled specific probes which have been described so far are not sensitive enough to be used for clinical speicimens without prior culturing. The advent of the polymerase chain reaction (PCR) has opened new possibilities for diagnosis of microbial infections. This technique has already been applied to a number of microorganisms. In the field of mycobacteria the PCR has been used to identify and to detect DNAs extracted from various mycobacteria. However, despite the extraordinary enthusiasm surrounding this technique and the considerable investiment, PCR has not emerged from the developmental "trenches" in the passed several years. It may be a considerable lenth of time before clinical microbiology laboratories become PCR playgrounds because many details remain to be worked out.

• Parasites, Hosts and Diseases
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• 제21권1호
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• pp.1-5
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• 1983

간츱충증 환자로부터 혈액을 채취하여 그 혈청에서 간흡충에 대한 항체를 Thin Layer Immunoassay(TIA)에 의하여 검출하였다. TIA는 Elwing et al.(1976)이 기술한 방법에 의하여 시행하였고, 항원은 이등(1981)의 방법으로 제조된 간흡충 성충 생리식염수 추출물을 사용하였으며 그 단백질 함량은 2.87mg/ml이었다. 간흡충중 환자 60명의 혈청에서의 항체가는 건강대조군 및 이질아메바 감염자에서의 항체가와 비교할 때 그 차이가 유의하였으나, 폐흡충증 환자와의 차이는 유의하지 않았다. 간흡충중 환자의 성별이나 연령과 TIA에 의한 항체가 사이에 상관 관계가 없었다. 환자의 분변내 EPG, 말초혈액내 호산구의 수와도 상관이 없었다. TIA에 의한 항체가와 간기능 검사 결과와의 관계를 보면 alkaline phosphatase, SGOT 및 SGPT 측정치와 모두 상관이 없음을 알 수 있었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권2호
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• pp.173-177
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• 2006

In this study, a novel strategy was developed for the highly selective immobilization of proteins, using the polyhydroxyalkanoate (PHA) depolymerase substrate binding domain (SBD) as an active binding domain. In order to determine the appropriacy of this method for immunodiagnostic assays, the single-chain antibody (ScFv) against the hepatitis B virus (HBV) preS2 surface protein and the severe acute respiratory syndrome coronavirus (SARS-CoV) envelope protein (SCVe) were fused to the SBD, then directly immobilized on PH A-coated slides via microspotting. The fluorescence-labeled HBV antigen and the antibody against SCVe were then utilized to examine specific interactions on the PHA-coated surfaces. Fluorescence signals were detected only at the spotted positions, thereby indicating a high degree of affinity and selectivity for their corresponding antigens/antibodies. Furthermore, we detected small amounts of ScFv-SBD (2.7 ng/mL) and SCVe-SBD fusion proteins (0.6ng/mL). Therefore, this microarray platform technology, using PHA and SBD, appears generally appropriate for immunodiagnosis, with no special requirements with regard to synthetic or chemical modification of the biomolecules or the solid surface.