• 제목/요약/키워드: indirect enzyme-linked immunosorbent assay

검색결과 99건 처리시간 0.026초

재조합 baculovirus에 의한 아프리카 돼지콜레라바이러스 p12 단백질의 발현과 진단적 적용 (Expression and diagnostic application of p12 protein of African swine fever virus by recombinant baculovirus)

  • 최강석;최정업;김용주
    • 대한수의학회지
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    • 제45권1호
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    • pp.63-70
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    • 2005
  • African swine fever (ASF) is an infectious disease of domestic and wild pigs for which there is no vaccine in the world. A proper surveillance of viral activity and a timely response to ASF outbreaks depend upon the rapid diagnosis of ASF viral infection. Internationally prescribed enzyme-linked immunosorbent assay (ELISA) is a fast, sensitive test routinely used in the diagnosis of the ASF. However, inactivated whole ASF virus antigen used in this test is a tedious to prepare and has a risk of outside exposure of infectious virus by laboratory accident during the preparation. An ASF virus noninfectious recombinant antigen is a safe and easily produced alternative antigen for use in diagnostic assay. We have cloned the ORF O61R gene of the ASF virus to generate a recombinant baculovirus producing the p12 protein in insect cells under control of the polyhedrin promoter as non-fusion protein. When used in an indirect ELISA, the p12 antigen showed reactivity with all known ASF positive pig sera but not with negative pig sera. Our results indicated that the p12 protein would be one of alternative antigens for diagnosis of the ASF.

Quantification of Immunoglobulin G against Trypanosoma cruzi in Individuals with Chronic Chagas Disease Treated with Nifurtimox and Evaluated in Prolonged Follow-Up

  • Munoz, Gabriela;Vergara, Camilo;Martinez, Gabriela;Apt, Werner;Zulantay, Ines
    • Parasites, Hosts and Diseases
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    • 제57권1호
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    • pp.39-41
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    • 2019
  • In the indeterminate chronic period of Chagas disease (ChD) the treatment has not been conclusive, because the serological negativization requires many years. This study aims to evaluate the efficacy of nifurtimox (NF) in the treatment of chronic ChD in prolonged follow-up by serological techniques of indirect immunofluorescence assay (IFA) and enzyme-linked immunosorbent assay (ELISA) IgG comparing 2 groups of patients, treated and non treated. Mann-Whitney test was performed for ELISA and IFA, with significant difference between the groups (P<0.05). IgG levels were lower in individuals treated compared with untreated patients, indicating chemotherapeutic efficacy in prolonged follow-up.

소 브루셀라병 표준시험관응집반응법 진단효율 평가 (Evaluation on diagnostic efficiency of the standard tube agglutination test for bovine brucellosis)

  • 성소라;김지연;허문;이기찬;구정희;강성일;이향근;김숙미;정석찬
    • 한국동물위생학회지
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    • 제35권4호
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    • pp.269-273
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    • 2012
  • A confirmatory serological test, the standard tube agglutination test (STAT) is evaluated for the diagnostic efficiency in brucellosis Korea. A total of 345 bovine samples were collected from regional veterinary branch under national brucellosis monitoring program from January 2010 to June 2012 in Korea. These samples were diagnosed as suspected serum and brucellosis positive by the Rose Bengal test (RBT) and the STAT, respectively. The STAT was compared and evaluated with three serological test such as the indirect-enzyme linked immunosorbent assay (I-ELISA), competitive-enzyme linked immunosorbent assay (C-ELISA) and fluorescence polarisation assay (FPA) prescribed for international trade by OIE. Among the 345 bovine serum samples, 302 (87.5%) were diagnosed as positive in the STAT, while 215 (62.3%), 223 (64.6%) and 194 (56.2%) serum samples were diagnosed as positive for brucellosis in the I-ELISA, C-ELISA and FPA, respectively. The STAT showed quite high positive results as compared with three prescribed tests of OIE. FPA, I-ELISA and C-ELISA have shown 60.6%, 64.9% and 67.2% correlation, respectively as compared to the STAT. However correlations of three prescribed tests ranged high 84.1~97.7%. Especially, correlation between I-ELISA and C-ELISA is quite high, 97.7%. These results suggest that the STAT has shown many false-positive reactions. Therefore, additional serological test, such as ELISAs and FPA, would be necessary to adopt as a confirmatory test in the national surveillance program of bovine brucellosis in Korea.

Metalaxyl의 다클론 항체생산과 최적 ELISA 조건 (Production of Polycolonal Antibody and Optimum Conditions in ELISA for Metalaxyl)

  • 이강봉;임건재;정영호;서용택
    • 한국환경농학회지
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    • 제13권1호
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    • pp.76-82
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    • 1994
  • Metalaxyl을 ELISA기법을 이용하여 분석하기 위하여 metalaxyl-HSA를 합성하고 이에 대한 항체를 생산하여 competitive indirect ELISA를 위한 최적조건을 조사하였다. Metalaxyl-protein conjugate는 metalaxyl을 가수분해시켜 생성된 metalaxyl acid에 단백질(HSA, OA)을 mixed anhydride방법이나 EDC 첨가방법으로 조제하였고 생성된 항체의 역가는 1:16,000으로 나타났다. Coating Ag의 최적농도는 $8\;{\mu}g/ml$, 배양시간은 $4\;^{\circ}C$에서 1시간 이상, $20\;^{\circ}C$$37\;^{\circ}C$에서는 4시간으로 나타났으며 항체의 희석배수는 1:2,000이 최적으로 나타났다. 항원과 항체의 혼합비율은 0.5 ml의 항원에 대해 1 ml의 항체가 최적이었으며 이 혼합액의 배양시간은 1시간, antirabbit Ig G-peroxidase conjugate의 배양시간은 30분이 최적으로 나타났다.

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신증후출혈열 환자의 혈청학적 및 분자생물학적 진단 검사법 비교 (Comparative Diagnostic Studies on Serologic and Molecular Biological Tests Against Haemorrhagic Fever with Renal Syndrome)

  • 우영대;문희주;배형준
    • 대한의생명과학회지
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    • 제6권2호
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    • pp.141-149
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    • 2000
  • 우리 나라에서 발생하고 있는 급성 출혈성 질환인 신증후출혈열의 원인 바이러스는 Family Bunyaviridae의 Genus Hantavirus에 속하는 한탄과 서울바이러스에 의하여 발생되고 있다. 본 연구에서는 신증후출혈열로 의뢰된 환자에서 한탄바이러스에 대한 항체가를 간접면역형광항체법(indirect immunofluorescent antibody technique, IFAT), 면역효소측정법 (enzyme-linked immunosorbent assay, ELISA) (IgG, IgM), 고비중입자응집반응 (high density composite particle agglutination, HDPA) 및 플라크감소중화시험 (plaque reduction neutralization test, PRNT) 등으로 비교 측정하였고, 신증후출혈열 환자로 확진된 15명의 한탄바이러스 혈청형을 PRNT와 혈청형 특이 역전사 효소 중합효소연쇄반응(nested reverse transcriptase polymerase chain reaction, nested RT-PCR)으로 확인하였다. 신증후출혈열로 의뢰된 환자에서의 한탄바이러스에 대한 IFAT, ELISA (IgG, IgM), HDPA그리고 PRNT비교에서 형광항체, ELISA IgG,응집항체 및 중화항체는 8명 모두 높게 나타났으며, ELISA IgM은 5명에서는 현저히 높은 항체를 보유하고 있었다. 신증후출혈 열 환자 15명에서는 높은 형광항체와 중화항체 역가를 나타내었고, 15명 중 12명은 한탄바이러스, 2명은 서울바이러스에 대한 높은 중화항체를 갖고 있었으며, 1명은 두 바이러스에 대하여 동일한 항체 역가를 나타내었으며, 혈청형 특이 primer를 사용한 nested RT-PCR에서는 15명 중 3명과 1명만이 한탄바이러스와 서울바이러스 primer에 대해 RNA가 검출되었다.

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말라리아 진단시 재조합 Circumsporozoite 단백질의 유용성 평가 (The Evaluation of Recornbinant Circumsporozoite Protein in Malaria Diagnosis)

  • 이형우;이종수;이원자;조신형;이호자
    • 미생물학회지
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    • 제36권2호
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    • pp.142-149
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    • 2000
  • 국내에서 유행하는 Plasmodium vivax의 유전자를 조작하여 만든 재조항 Circumsporozrozite(CS) 단백질 (22)을 이용하여 말라리아에 대한 혈청학적 검사의 유용성을 평가하였다. 최초발병일로부터 진단까지 걸린 기간을 기준으로 환자들의 면역반응을 Western blot으로 알아본 결과, 15일 이내에 진단받은 환자들은 43.8%(14/32)가 양성반응을 보였고, 16일 이상 경과한 환자들은 94.4%(17/18)가 양성반응을 보여 전체적으로는 62%(31/50)의 양성률을 보였다. Blood stage 항원에 대한 항체를 갖고 있음에도 불구하고 재조합 CS단백질 항원에 대해 음성인 환자들이 22.6%(7/31)였다. 비유행지 주민(경북 예천군)들은 10.7%(3/28),유행시 주민(인천시 강화군)들은 27.6%(13/47)의 재조합 CS 단백질에 대한 항체 양성을 보였다. 재조합 CS 단백질 항원의 역학조사시 유용성을 알아보기 위하여 말라리아 유행지인 경기도 파주시 조산리, 마정리, 항양리, 뇌조리 거주 주민 422명의 전혈을 채취하여 혈액도말법과 중합효소연쇄 반응법으로 항원검사를 실시하였으며 blood stage 항원을 이용한 간접면역형광법과 재조합 CS 단백질 항원을 이용한 효소면역측정법으로 말라리아 항체보유 여부를 조사하였다. 혈액도말법에서는 422명 모두 음성이었으나, 중합효소연쇄반응법에서는 2명(0.47%)이 양성이었다. 간접면역형광법에서는 42명(9.95%), 재조합 CS 단백질 항원을 이용한 효소변역측정법에서는 71명(16.82%)이 양성으로 나타났다. 두 검사에서 모두 항체 양성을 보인 사람은 8명(11.27%)에 불과하였다.

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인천지역에서 발생한 돼지콜레라의 역학적 특성 (Characteristics of outbreak for the classical swine fever (CSF) at Incheon metropolitan area in 2002)

  • 권효정;변재원;이정구;김경호;박은정;이성모;황현순
    • 한국동물위생학회지
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    • 제26권1호
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    • pp.1-9
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    • 2003
  • The aim of this survey was to investigate the characteristics of outbreak farm determined as the classical swine fever(CSF) at Gangwha-gun and Seo-gu, Incheon metropolitan area from October 7 to November 25 in 2002. Sixty pigs in six different farms were confirmed to the CSF and a total of 9,106 pigs containing 3,194 related epidemiologically was slaughtered to stop spreading of the disease. Clinical signs of pigs diagnosed with the CSF were high fever, anorexia, depression, paralysis of hindlimbs, cyanosis, etc and gross lesions were typically represented with hemorrhage of submandibular and superficial lymph node, infarction of spleen, and petechial (ecchymotic) hemorrhage of kidney and skin. But some outbreak farms had not shown remarkable symptoms, so they were confused with other bacterial diseases. White blood cell (WBC) counts, the classical swine fever virus(CSFV) antigen and antibody enzyme linked immunosorbent assay(ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR) results about six farms indicated that total 60 pigs were infected with the CSFV. Although the origin and infection route of the CSFV were not clear, but the transmissions between farms were mainly through indirect contact such as the movement of farm personal and vehicles from outbreak farm.

Development of an ELISA for the Detection of Fenazaquin Residues in Fruits

  • Lee, Jae-Koo;Kim, Yun-Jung;Lee, Eun-Young;Kim, Dae-Kyu;Kyung, Kee-Sung
    • Journal of Applied Biological Chemistry
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    • 제48권1호
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    • pp.16-25
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    • 2005
  • To develop an enzyme-linked immunosorbent assay (ELISA) for the detection of the residues of the acaricide fenazaquin, five haptens were synthesized and assessed. A competitive indirect format was used with polyclonal antibodies. Under an optimized condition using the selected rabbit C antiserum, an $IC_{50}$ of $96.97\;ng{\cdot}ml^{-1}$, the detection range of $14.9{\sim}631\;ng{\cdot}ml^{-1}$, and the lowest detection limit of $8\;ng{\cdot}ml^{-1}$ were obtained. Some structurally related compounds of practical use showed low crossreactivities to the antibody. Highest cross-reactivity observed with hapten IV indicates that the antiserum C recognizes very well quinazoline ring, 4-tert-butylphenyl, and an adequate length of spacer arm. The length of spacer arm affected recognition of quinazoline ring and 4-tert-butylphenyl moieties. When applied to apple and pear, recoveries were within acceptable ranges of $93.18{\sim}104.77%$ (n = 4) and $79.40{\sim}111.95%$ (n = 4), respectively.

감마선 조사된 우유단백질에 대한 우유 알러지 환자의 IgE 결합능의 변화 (Changes of Binding Ability of Milk-Hypersensitive Patients질 IgE to Gamma-Irradiated Milk Proteins)

  • 조경환;육홍선;이주운;이수영;변명우
    • 한국식품영양과학회지
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    • 제30권3호
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    • pp.505-509
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    • 2001
  • This study was carried out to evaluate the application of food irradiation technology as a method for reducing milk allergies. Bovine $\alpha$-casein, $\beta$-casein, $textsc{k}$-casein, $\alpha$-lactalbumin(ALA), $\beta$-lactoglobulin (BLG) and serum albumin (BSA) were used as model allergens of milk proteins and the proten solution (2.0 mg/mL) with 0.01 M phosphate buffered saline (pH 7.4) was irradiated at 3, 5 and 10 kGy. Using milk-hypersensitive patients IgE (MHP-IgE), the changes of binding ability to irradiated proteins were observed by competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA). Affinity of MHP-IgE to milk proteins was higher in ALA and BLG than that of other proteins. Standard curve to each non-irradiated protein could be made with MHP-IgE for quantifying milk allergens. Binding abilities of MHP-IgE to the irradiated proteins, however, decreased with different slopes of the standard curves. Sensitivity of gamma irradiation was higher in ALA and BLG than of other proteins. These results indicated that irradiation technology can be used to reduce the milk hypersensitivity.

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대전지역 길고양이의 톡소포자충(Toxoplasma gondii) 감염 실태 조사 (Investigation of Toxoplasma gondii infection on stray cats in Daejeon)

  • 성선혜;유상식;임여정;정년기;문병천
    • 한국동물위생학회지
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    • 제35권1호
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    • pp.19-24
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    • 2012
  • This study was performed to evaluate the prevalence rate of Toxoplasma gondii on 217 stray cats in Daejeon. The positive infection rate of T. gondii was 15.7% in enzyme-linked immunosorbent assay (ELISA), 12.4% in latex agglutination test (LAT), 14.7% in indirect immunofluorescent antibody test (IFA) and 0.5% in polymerase chain reaction (PCR) respectively. In districts, Yuseong-gu was shown the highest seropositive rate of T. gondii as 31.8% in ELISA, 22.7% in LAT and 31.8% in IFA. In gender, the seropositive rate of female cats was slightly higher than that of male cats as 17.2% in ELISA, 15.2% in LAT, 15.2% in IFA and 1.0% in PCR. Cats captured in National science museum, detached house and apartment was shown relatively high prevalence rate of T. gondii.