• 제목/요약/키워드: antiserum

검색결과 296건 처리시간 0.027초

효소면역 전기영동 이적법을 이용한 질트리코모나스 항원의 비교 분석 (Comparative antigen analysis of yrichomonus vaginulis by enzyme-linked immunoelectrotransfer blot technique)

  • 민득영;임미혜
    • Parasites, Hosts and Diseases
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    • 제30권4호
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    • pp.323-328
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    • 1992
  • 질트리코모나스의 항원 분석을 시행하기 위하여 병원성이 화인된 질트리코모나스(Trichamonas vaginnlis) 6개주(HY-1,2,3,9,10,13)의 항일을 sodium dodecyl sulfate - polyacrlyamide gel electrophoresis(SDS-PAGE) 한 결과 Coomassie brilliant blue 염색상에서 peptide의 밀도 차이를 제외하고는 주간에 동일한 단백질 분포양상을 보였으며, 12kDa에서 170kDa까지 약 35개 정도의 분획이 관찰되었다. 또한 질트리코모나스 HY월주를 항원으로 하고 질트리코모나스 6개주에 대해 면역된 마우스의 항혈청을 이용하여 enzyme-linked immunoelectrotransfer blot(EITB)을 시행한 결과 각 주마다 서로 다른 반응양상을 보였으며, 51kDa과 96kDa에서 질트리코모나스의 특이한 공통 반응대가 관찰되었고, 각기 다른 6개주의 항원에 대해 HY-1주의 항혈청으로 ETTB한 결과 HY-1 함원(homologous antigen)과의 반응 양상과 타항친(heterologous antigen)과의 반응 양상간의 특이한 차이는 관찰할 수 없었다. 이상의 결과로 보아 질트리코모나스 각 주의 항원은 항인-항체 반응에서 주간에 이종(antigenic heterogeneity)을 형성하고 있는 것으로 보였으며, 41, 47, 55, 74 및 94kDa에서 질트리코모나스애 특이한 공통반응대를 보었으며, 이 부분이 숙주-기생충의 상 호관계에 있어서 중요한 의의를 내포하고 있을 것으로 생각된다.

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돼지 분변 유래 PORCINE ENTEROVIRUS의 물리화학적 특성 및 병원성에 관한 연구 (Studies on Physico-chemical Properties an d Pathogenicity of Porcine Enterovirus Isolated from Feces of Pigherds)

  • 박정우;이종인;신용호;조우영;최윤식
    • 한국동물위생학회지
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    • 제14권2호
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    • pp.110-120
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    • 1991
  • 28 porcine enteroviruses were isolated from 86 pig-feces of 9 swine farms located in south region, Chung-buk, from March to September 1990. Physico-chemical properties and pathogenicity of isolates were investigated. Results obtained throughout experiments are summarized as follows. According to the age, weanlings(40-90 days), sucklings(10-30 days) and adult pigs(6 months over) showed the isolation rate of 67%. 8% and 4%, respectively. By physico-chemical tests, YD-90/22, YD-90/43 and YD-90/64 strains were found to be ether, chloroform and PH stable. Nucleic acid test suggests the virus to have a DNA genome. Most of the Isolates were not evident of hemagglutinin using erythrocytes from various mammalian & avian. 22 strains among the isolates were shown CPE type I and the remainders were CPE type II. 3 strains among isolates of CPE type I strains were neutralized with high titers to serotype 2 antiserum. In the study on virus growth curve in PK-l5 cells, YD-90/22, YD-90/43 and YD-90/64 strains showed the maximum infectivity titers($10^{6.0}-l0^{6.5} TCID({50}ml$) at 4days post inoculation(PI). When 30 day-old commercial piglets were inoculated only intraoral route with the YD-90/22 strain at $10^{6.0} TCID_{50}ml,$ piglets not showed the symptoms. But piglets inoculated by intramuscle route, intraoral and intramuscle route after pretreat with dexamethasone(2.5mg /kg) for 5 days were shown the symptoms of anorexia, diarrhea, pyrexia and ataxia at 4th-6th days PI. The viral reisolation in the virus-inoculated piglets was examined from feces. The viruses were recovered intermittently from 2nd to 16th day PI and at 4th-6th day PI, all piglets excreted viruses.

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Characteristics of ustilago maydis virus of SH14 killer strain isolated in Korea

  • Hwang, Seon-Hee;Jung, Cheong-Hwan;Yie, Se-Won
    • Journal of Microbiology
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    • 제33권2호
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    • pp.154-159
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    • 1995
  • SH-14, a novel killer strain of Ustilago maydis was isolated in Korea. It has been reported in other papers that the toxin specificity and double-stranded RNA pattern of SH-14 strain were different from other laboratory strains. In this paper, we analyzed the biochemical characteristics of U. maydis SH-14 virus. Three distinctive peaks were isolated from CsCl density gradient, designated as top (T), intermediate (I) and bottom (B) components. We found that the densities of each components, 1.285, 1.408 g/cm$\^$3/, respectively, are very similar to those of other strains. As previously reported by the analysis of dsRNA in each component, the dsRNA segments are separately encapsidated. Capsid protein of SH-14 virus consists of two proteins about 70 Kd shown by SDS-PAGE analysis. Electron microscopic examination of the virus particles revealed that UmV particles are very similar in size and morphology to all isolates as well as all lab-strains. In order to test immunological cross reactivity of UmV, werstern bolt analysis was carriedout with antiserum against A8 virus. All capsid protein had positive reaction against A8 antibody which indicated that UmV are immunologically cross-reactive with all isolates from Korea. The results presented in this paper may show that UmV isolated from SH-14 strain has very similar biochemical characteristics to those of other UmV. However, the difference in the toxin specificity and the molecular weight of toxin protein from the SH-14 strain has us to conclude that U. maydis SH-14 strain is a new killer type.

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가잠에서 분리된 새로운 미포자충 K79의 병리학적 연구 I. 미포자충 포자의 정제 및 식별 (Pathological Studies on the New Microsporidia K79 Isolated from the Silkworm, Bombyx mori L. I. Purification and Serological Discrimination of Microsporidian Spores.)

  • 윤재수;임종성
    • 한국잠사곤충학회지
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    • 제26권2호
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    • pp.1-6
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    • 1984
  • 미포자충 포자의 정제법을 개발하고 포자표면의 혈청학적 반응을 구명하기 위하여 수행한 시험결과를 요약하면 다음과 같다. 1. 미포자충 포자의 분리정제는 예비원심-percoll 평형밀도원심-세척의 3단계법이 우수하였고, 분리정제된 포자는 극사추출에 나쁜 영향도 받지 않았다. 2. 포자의 형태는 K79 포자가 장타원형(장단경비 2.76)이고 N. bombycis는 단타원형(장단경비 2.07)으로 양포자가 상이하였다. 3. 형광항체 간접법에 의하여 항 N. bombycis 포자혈청에 대하여는 N. bomvycis 포자만이 강한 형광을 발현하였으나 항 K79 포자의 혈청에 대하여는 10배희석구에서는 양포자 모두 강하게, 20배 이상 희석구에서는 약하게 형광을 발현하였고 K79 포자의 특이항체가 포자외각에 인정되었다.

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갑상선질환(甲狀腺疾患)의 Thyroglobulin에 관(關)한 연구(硏究)(예보(豫報)) (The Study on the Thyroglobulin in Thyroid Diseases)

  • 김석근;임수덕;이문호
    • 대한핵의학회지
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    • 제4권1호
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    • pp.43-49
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    • 1970
  • In recent years with development of immuno-electrophoresis, more acurate analysis of the serum protein became possible. However, there is few reports in the literature which investigated the changes of the immunoglobulin compared with electrophoretically fractioned serum thyroglobulin in the patients with various thyroid diseases. The purpose of this report is to investigate the changes of thyroglobulin in various thyroid diseases by the method of immuno-electrophoresis and to compare the results with.serum protein fractionated by the method of agar-gel micro-electrophoresis. Materials and Methods: Sera from 9 patients with diffuse toxic goiter, 2 nodular nontoxic goiter, 2 thyroiditis, 3 hypothy, roidism, 1 thyroid cancer, 7 cystic degeneration of the thyroid gland, and 10 normal subject were taken. All cases were confirmed by various laboratory thyroid function tests and thyroid needle biopsy. Immuno-electrophoretic analysis of the serum were performed by Scheidegger's modified micro-immuno-electrophoretic method. The antiserum was obtained from the Travenol Laboratories International, Hyland Products Division and was rabbit anti-human thyroglobulin. Microscope slide agar-gel electrophoresis for serum protein fractionation was performed at $4^{\circ}C$ using veronal buffer, pH 8.6 and ionic strength 0.05, with 54 volts and 2.8 mA for 60 minutes. The fractionated slide was stained with 0.1% thiazine red. The results were as follows: 1) Increase of immune-globulin macroglobulin (IgM), alphaglobulin, and immune-globulin A (IgA) by 95.8%, 100%, 29.2% respectively was found in the serum from various thyroid diseases. 2) Thyroglobulin fraction was found to be increased in 50%, no change in 41.7%, and no line in 8.3% with all of the various goiter patients. On the other hand, 10 normal control group showed only 2 cases of increase, 5 cases of no change and 3 cases of no line.

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Progesterone 측정을 위한 면역분석법의 최적조건에 관한 연구 (Optimization of Immunosasay Procedures for the measurement of Progesterone)

  • 강원준;고대환;이경광;김종배;정길생
    • 한국가축번식학회지
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    • 제9권2호
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    • pp.105-112
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    • 1985
  • This research was carried out to investigate the optimal conditions ssociated with the RIA procedures such as a bridging phenomena, prozone effects and a new separation methods etc. The results obtained were summarized as follows: 1. The lgG fractions of donkey anti-rabbit IgG sera were purified by Protein-A-Sepharose affinity column, which indicates that Protein-A an affinity for IgG class of donkey antiserum. 2. In coating the IgG fraction on polystryene tubes, incubation conditions made no differences between 2 hr at room temperature and overnight at 4$^{\circ}C$. 3. There were no significant differences between 1st antibody-coated tube and 2nd antibody-coated tube as a separation method when compared in terms of reproducbility. A better reproducibility may be expected if the titers of 1st antibody for the progesterone to be assayed and of corresponding 2nd antibody are reasonably high. 4. The titers of anti-progesterone antibody for 3H-progesterone and progesterone-11HS-125I were 1:300 and 1:700 in liquid-phase, and 1:100 and 1:300 in solid-phse for the separation methods. 5. A bridging phenomena in which a standard curve is long and shallow were observed when progesterone-11HS-125I was used for the tracer, but not in 3H-progesterone. 6. A prozone effect in a solid-phase system, especially 1st antibody-coated tube method was observed which the degree of inhibition was significantly different although zero bindings look the same. In this case, the titration curve should be made both in the absence and in the presence of a, pp.opriate amount of competiter, standard, respectively.

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한국 울릉도 근해의 비브리오속의 분리연구 (Studies on the Isolation of Vibrio Species from Sea Water and Marine Products at Ulleung Island of Korea in 1985)

  • 주진우;이미헌;김일
    • 대한미생물학회지
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    • 제21권3호
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    • pp.345-353
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    • 1986
  • Authors studies on the isolation of non-sucrose fermentation Vibrio species such as V. parahaemolyticus, V. vulnificus and V. damsela from sea water, shellfishes and various algae at Ulleung island in the east of Korea on summer in 1985. Authors carried out test for isolated strains to biochemical characteristics, halophilism, hemolysis and serological reaction. The results obtained were as follows: 1. Strains isolated from total specimens were 25 strains of V. parahaemolyticus, 2 strains of V. vulnificus and 1 strain of V. damsela, respectively. 2. Isolated frequency of Vibrio species from sea water and marine products was lower than on specimens in the southern sea of Korea from 1980 to 1985. 3. On hemolysis reaction on blood agar media using human and rabbit erythrocytes, 14 strains among 25 strains of V. parahaemolyticus isolated were positive, and V. vulnificus and V. damsela were positive, respectively. 4. The distributions of serotypes of V. parahaemolyticus isolated were from O1 group to O10 group, and from K 10 to K 69. The results of K serotypes were as follows ; serotype K 10 and K 33 were 2 strains, serotype K 59 and K 69 were 1 strain, respectively. And 2 strains of V. vulnificus isolated were not agglutinated by antiserum of V. vulnificus. Therefore, the probability is that 2 strains of V. vulnificus isolated were other serotypes distributed on sea weater and marine products of Ulleung island of Korea.

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Identification and Cloning of a Fraction 1 Protein of Yersinia pestis that Produces Protective Immune Responses

  • Kim Jong-Hyun;Cho Seung-Hak;Jang Hyun-Chul;Lee Hee-Cheul;Kim Young-Il;Kang Yeon-Ho;Lee Bok-Kwon
    • 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.

Molecular Clonging and Hyperexpression of a Bt Gene, cryIAc, in Escherichia coli $DH5{\alpha}$: Production and Usage of Anti-CryIAc Antibody

  • RYOU, CHONGSUK;TAEYOUNG CHUNG;MOOSIK KWON
    • Journal of Microbiology and Biotechnology
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    • 제11권6호
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    • pp.1093-1098
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    • 2001
  • The gene coding for a Lepidoptera-specific insecticidal crystalline (or control) protein (ICP), recognized as cryIAc, from Bacillus thuringiensis subsp. kurstaki HD-73, was cloned into the vector pBluscript ll SK-, and then transformed in Escherichia coli $DH5{\alpha}$. The clone was named EBtIAc and the chimeric phagemid, as pEBtIAc. Hyperexpression of CryIAc protoxin was observed in the extract of the culture of E. coli harboring pEBtIAc. Crystalline protoxin was purified by differential solubility. It was dissolved in alkaline pH, and exposed to trypsin to be activated. The molecular weights of the pro- and activated toxins on SDS-PAGE were estimated to be ca. 130 kDa and 60 kDa, respectively. The toxicity was tested by force-feeding larvae of gypsi moth (Lymantria diapar) with trypsinized protoxin. Using the batch of biologically active form of the toxin as an immunogen, anti-CryIAc antiserum was raised in a New Zealand white rabbit. Immunoglobulin G was fractionated from the seam by Protein-A sepharose affinity chromatography. Immunoreactivity of the antibody was examined by dot and Westerns blottings. It has been found that the anti- CryIAc antibody recognized the purified toxin at a level below a nanogram in terms of quantity. Using the antibody some of Bt-corns were able to be differentiated from tons of corn kernels which were imported from America as forage crops.

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Monitoring Ovarian Function by Solid- Phase Chemiluminescence Immunoassay

  • 김종배;구병삼
    • Clinical and Experimental Reproductive Medicine
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    • 제9권1_2호
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    • pp.43-53
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    • 1982
  • 여성의 난소기능은 뇨중 Oestrone-3-glucuronide를 간편한 solid-phase 의 화학발광성 면역학적 측정법 (Chemiluminescence Imrnunoassay(CIA) 에 의하여 그 기능이 탐지될 수 있다. Oestrone-3-g1ucuronyl-6-bovine serum albumine에 대한 antiserum의 IgG fraction은 polystyrene 실험관벽에 흡착시켰으며, 항원으로서는 est r one-3- gl ucuronyI-6-aminoethyl-ethyl-isoluminol 을 항원 (antigen) 에 labeI 시킨 것이다. 시험 대상물인 뇨는 매일아침뇨(early morning urine) 을 희석 (1:1000 V/V)한 후 100mcl 를 취하여 이를 각기 이중분석액으로 택하였다. 시험관 내에서 결합반응 (1 hour at $4^{\circ}C)이 일어난 후에는 시험관내의 액체를 전부 흡입 폐기시켰으며, 항체반응이 일어난 후 ( antibody-bound fraction )에는 완충액 (400mcl)으로 한번 세척시켰다. 그후 염화수산화물(2N , 200mcl)을 가지고 $22^{\circ}C$에 60 분간 방치 혼합케 한 후 효소(microperoxidase) 와 과산화수소를 가하면서 산화작용에서 발생되는 발광양을 10초동안 측정하여 그 결과를 분석하였다. 위에 기술한 분석방법을 평가하면 다음과 같은 결론을 얻었다. Calibration curve sensitivity$3.12{\pm}0.75$ PG/tube ($mean{\pm}SD$)였고, lntra-assay precision(CV%) 9.52 (20 replicates;$38.4{\pm}3.66$nmol/1) 와 8.81 (15 replicates; $102.4{\pm}8.82$nmol/1)였다. Inter-assay precision(CV%) 은 11.9 (mean of 4 pools-7.03, 23.16, 52.11 과 117.53 nmol/1)로 2개월 동안에 걸쳐 시행되었고, 평균 비이어스(mean bias)는 -0.78 로 28에서 448 nmol 범위로서 매일아침 "뇨"의 차이분(different aliquots)은 좋은 결과를 얻었다. 건강한 여성으로부터 채취된 뇨중 Oestrone-3-glucuronide 의 농도(nmol/1)를 보면 월경주기의 여포기와 배난기 및 황체기에 있어서 각기 $40.2{\pm}9.9$ , $102.3{\pm}39.4$$84.3{\pm}13.3$nmol/1였다. 이와같은 결과는 동일한 검사뇨를 방사면역학적 방법(RIA)으로 측정 (6 menstrual cycle)한 결과와 유사한 측정치를 얻으므로서 간편하고 진보된 좋은 방법중의 하나라고 사료되는바이다.

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