• 제목/요약/키워드: indirect-ELISA

검색결과 178건 처리시간 0.025초

ELISA법에 의한 mouse의 혈청 및 조직중의 T-2 toxin의 검색 (The Detection of T-2 toxin in Serum and Organ of Mouse by ELISA)

  • 김동술;송재영;정덕화
    • 한국환경보건학회지
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    • 제22권1호
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    • pp.51-56
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    • 1996
  • In order to detect the T-2 toxin accumulation in the animal tissues, T-2 toxin, produced by Fusarium sporotrichioides M-1-1, was injected to mouse by 0, 1 and 2 mg per kilogram of body weight, respectively, and T-2 toxin extracted from serum and organs were analyzed by the indirected competitive ELISA. The indirect competitive ELISA established in the laboratory can be check less than 0.1 ppb level of T-2 toxin and average recovery of T-2 toxin spiked was 80~113% in animal samples such as serum, liver and kidney. After 6 weeks of treatment with 2 mg of T-2 toxin per kg body weight, T-2 toxin was accumulated in serum (133.0 ng/ml), liver(1.4 ng/g) and kidney(14.3 ng/g) of mouse injected with 2 mg of toxin per kg body weight.

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간접 Latex 응집반응과 ELISA에 의한 중추신경계 질환 환자의 혈청 및 뇌척수액에서 Toxoplasmu gondii에 대한 항체 검출 (Detection of Antibodies in Serum and Cerebrospinal Fluid to Tonoplasma gondii by Indirect Latex Agglutination Test and Enzyme-Linked Immunosorbent Assay)

  • 최원영;남호우
    • Parasites, Hosts and Diseases
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    • 제30권2호
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    • pp.83-90
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    • 1992
  • Toxoplasma증의 혈청학적 진단에 있어서 민감도를 증가시키기 위해 간접 latex 응집반응의 결과와 비교하면서 ELISA를 개발하였으며, 뇌척수액의 검사 시료로서의 가능성을 검토하였다. 아울러 중추신경계 질환환자로부 터기생충질환을 감별하기 위하여 1986년부터 1991년까지 전국 카 병원에서 채취한 혈청과 뇌척수액에 대하여 간접 latex 응집반응(ILA)과 ELISA를 실시하여 Toxoplasma 항체 보유 양상을 비교 검토하였다. 전체 2,016 건의 혈청에 대해 ILA를 실시하여 76건(3.8%)의 양성 (1:32이상의 titer)을 얻었다. 그러나 양성 혈청환자에서 채취한 뇌척수액에서는 낮은 titer의 반응은 있었으나 양성은 나타나지 않았다. 이들 양성 혈청의 양성 혈청 및 음성 혈청에 대하여 ELISA로 항체검사를 실시한바 ILA의 titer가 1 : 32인 군에서 통계적으로 유의한 차이를 나타내는 항체값을 얻었으며, 그 흡반도는 0.40이었다. 뇌척수액에 대한 ELISA로는 ILA의 1 : 64 titer군에서 통계 적으로 유의한 차이가 나타났고 그때의 흡광도 0.27을 양성 판단의 기준으로 사용하였다. ELISA에 의한 항체 검사상 전체 혈청에서 7.0%의 양성을 검출하여 ILA보다 약 2배 정도의 높은 민감도를 보였으며, 뇌척수액에서 는 5.6%의 양성률을 보여 ELISA는 뇌척수액에서의 항체 검출시 유용한 방법이라고 판단하였다. ILA에 비하여 ELISA는 약 2배 정도 높은 양성률을 내었고 양성률은 나이에 마라 40대 이후 급격한 증가를 보였으며, 여성보 다는 남성에서 약 2배 정도 양성률이 높게 나타났다. ELISA에 의한 뇌척수액의 항체 검사에서는 양성률의 성별 차이를 나타내지 않았다. 이상의 결과로 판단할 때, ELISA가 ILA보다 Toxoplasma 항체 검출의 민감도가 높았으며, 뇌척수액은 ELISA의 좋은 검사시료가 되며, 특히 중추신경계 Tocxoplnsma증의 진단에 있어 뇌척수액에 대한 항체 검사에서 ELISA가 유용하다고 판단하였다.

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돼지 생식기호흡기증후군 바이러스항체 검색에 있어 간접형광항체법(IFA) 과 효소면역법(ELISA)의 진단효율 비교 (Comparison between indirect immunofluorescent antibody (IFA) test and enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to porcine reproductive and respiratory syndrome virus(PRRSV))

  • 박최규;류영수;이창희;정종욱
    • 대한수의학회지
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    • 제38권2호
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    • pp.314-318
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    • 1998
  • An establishment of effective control measures to PRRSV infection in swine industry depends on a sensitive and specific diagnosis to detect either viral antigen and/or antibodies to PRRSV. Several diagnostic methods are available to detect antibodies against PRRSV, including IPMA, IFA and ELISA tests have been successfully developed. Sensitivity of the indirect immunofluorescent assay in MA-104 cells using Korean field isolate PL96-1 was superior to that of VR-2332 and field isolate PL96-2. Sensitivity and specificity of the IFA test with PL96-1 were comparable to those of commercial ELISA test kit but ELISA test was more sensitive for the detection of declining antibodies to PRRSV in finishing pigs. In this study we concluded that IFA and ELISA test could be utilized to detect antibodies to PRRSV and the results generated from these two tests were comparable and there were no significant difference between these two tests.

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ELISA 및 간접 latex 응집반응검사에 의한 임산부의 항 톡소포자충 항체가 (Toxoplasma antibody titers by ELISA and indirect latex agglutination test in pregnant women)

  • 류재숙;민득영
    • Parasites, Hosts and Diseases
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    • 제34권4호
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    • pp.233-238
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    • 1996
  • 이 연구에서는 경기도 양평군 및 광주군에 사는 임산부 899명을 대상으로 IgG-ELISA와 간접 latex 응집반응검사를 시행하여 톡소포자충에 대한 항체가를 측정하였다. IgG-ELISA에서는 0.25 이상을 양성기준으로 하였을 때 음성대조군 218명 중 4명이 양성(1.8%)인 반면 임산부에서는 39 명이 양성으로 검출되어 4.3%의 양성율을 보였다. 간접 latex 응집반응검사는 수의과학연구소에서 만든 킷트(LAT)를 사용하였는데 1:64 희석배수 이상을 양성으로 하였을 때 음성대조군은 모두 음성반응을 보였고 임산부에서는 7명(0.8%)이 양성을 보였다. 임산부중에서 1.8 이상의 반응을 보인 80명을 대상으로 일본제품인 Toxotest-MF를 적용시키고 1:32 이상을 양성의 기준으로 하였을 때 임산부 8명에서 양성반응을 보였다. LAT와 Toxotest-MT의 두 반응간의 일치율은 0.94(${\kappa}-index$ = 0.632. p < 0.01)로 높은 일치율(fair to good agreement)을 보였으므로 LAT는 톡소포자충증의 예비진단에 이용될 수 있을 것으로 생각된다.

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Early Detection of Epiphytic Anthracnose Inoculum on Phyllosphere of Diospyros kaki var. domestica

  • Lee, Jung-Han;Han, Ki-Soo;Lee, Sun-Cheol;Shim, Chang-Ki;Bae, Dong-Won;Kim, Dong-Kil;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • 제20권4호
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    • pp.247-251
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    • 2004
  • We developed a polyclonal antibody (PAh) based- ELISA system to accurately and rapidly monitor inocula on plant surface before onset of anthracnose. Titer of mouse antisera against conidia of Colletotrichum gloeosporioides was determined by using indirect ELISA. It was high enough to be detectable up to ${\times}$ 12,800 dilutions. Absorbance readings exceeded (1.5even at a 10$^{-5}$ dilution. Sensitivity of PAb was precise enough to detect spore concentration as low as 50 conidia/well by indirect ELISA. PAb1 and PAb2 proved to be very sensitive and highly specific to the target pathogen, C. gloeosporioides, apparently discriminating other unrelated pathogens, or epiphytes. Absorbance values for original isolate exceeded 1.0, but no reaction was detected with other isolates, except three other anthracnose fungi: C. gloeosporioides (pepper strain), Glomerella cingulata (apple strain) and C. lagenarium. Our data suggest that PAb1 and PAb2 bind with the protein epitope that partially contains residues of amino acid, arginine, and Iysine. This kit fulfills the require-ments for detecting inoculums before infection and during onset of anthracnose on sweet persimmon.

Analysis of Microtoxins in the Nakdong River Watershed

  • Park, Jung-Min;Lee, Jae-Jung;Hwang, Dong-Jin;Yang, Sang-Yong
    • 한국환경과학회:학술대회논문집
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    • 한국환경과학회 2003년도 International Symposium on Clean Environment
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    • pp.99-102
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    • 2003
  • The different methods such as HPLC, indirect- and direct-ELISA were employed for the analysis of microtoxins and the results of each method were compared in terms of the detection limit and accurary. Three toxins, microcystin-RR, -LR and -YR were clearly separated by HPLC using 0.05 M methanol and phosphate buffer used as a solvent system. The calibration curves for the toxins were linear in the range of 5 ng to 50 ng. The standard curves for the immunoassay of microcystin obtained by direct and indirect ELISA are compared. The linear responses of inhibitions of binding by microcystin in the direct and indirect competitive ELISA were in the range of 10 ng to 1000 ng and 50 pg to 160 pg, respectively. Distribution of microtoxins at 11 sites in the Nakdong river and several lakes in Korea was also studied. The most dominant microcystin variant in the test sites was found to be microcystin-RR.

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IHA(Indirect hemagglutination test)를 이용한 간흡충증(肝吸虫症)의 면역혈청학적(免疫血淸學的) 진단(診斷)에 관한 연구(硏究) -ELISA법과의 비교(比較) 및 치료(治療) 전후(前後) 의 혈청항체가(血淸抗體價) 변동(變動)에 관하여- (Changes of Serum antibody Levels by Indirect Hemagglutination Test after Treatment in Clonorchiasis)

  • 주경환;김종성;정명숙;임한종
    • 농촌의학ㆍ지역보건
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    • 제12권1호
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    • pp.117-123
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    • 1987
  • Detection of IgG antibody in clonorchiasis has been accomplished through various serodiagnostic procedure including complement fixation test, gel diffusion test, indirect fluorescent antibody test, indirect hemagglutination test etc. In this report enzyme immunoassay (ELISA) and indirect hemagglutination test (IHA) were used to determine IgG serum antibody levels before and after therapy with praziquantel. Briefly, sera from 62 cases of confirmed human clonorchiasis were examined before and after treatment with praziquantel. Among 62 cases treated 25 cases were categorized as completely cured groups by formalin-ether and careful examination of 4 cellophane thick smered slides at 18 months after treatment. The sera of 25 cases of cured groups were examined again by ELISA and IHA, and com-pared to the previous data. The results obtained were as follows; 1) Sensitivity of IHA test was 83.6% when cut-off titer of 1:8 was applied. No sera obtained from 10 normal healthy control showed positive reaction. 2) Twenty cases (80.0%) out of 25 cured one showed negative results by IHA at 18 months after treatment. 3) Although 5 cases showed positive titer even 18 months after treatment 3 cases of them showed decreased antibody titer. However 2 cases did not show any response. 4) Even though almost all cases showed de- creased ELISA value, only 11 cases (44.0%) out of 25 patients showed negative results by ELISA at 18 months after treatment. In conclusion, it is suggested that, while IgG ELISA for detecting long persisting antibody was more sensitive than IHA, IHA results more conclusively indicated effective treatment in clonorchiasis by negative conversion than did the results of ELISA.

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Evaluation of Two ELISA and Two Indirect Hemagglutination Tests for Serodiagnosis of Pulmonary Hydatid Disease

  • Eris, Fatma Nur;Akisu, Ciler;Aksoy, Umit
    • Parasites, Hosts and Diseases
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    • 제47권4호
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    • pp.427-429
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    • 2009
  • To establish a definite diagnosis for pulmonary hydatid disease, combination of radiology and serology is useful. In this study, 19 preoperative sera from patients with surgically confirmed pulmonary hydatidosis, 40 sera from patients with other parasitosis and pulmonary diseases, and 20 sera from healthy donors were evaluated using 4 different serological tests, i.e., the commercial ELISA (ELISA-kit) test, the ELISA (ELISA-lab) test prepared in our laboratory, the commercial indirect hemagglutination assay kit (IHA-kit) test, and the IHA test using sensitized sheep red blood cells with tannic acid (IHA-TA). The ELISA-kit was the most sensitive (84.2%) and the most specific test (100.0%). The ELISA-kit also demonstrated the highest positive (100.0%) and negative (95.2%) predictive values. The sensitivity of the ELISA-lab test, that we prepared, was found to be 73.6%, whereas the IHA-kit test and the IHA-TA test were found to be 73.6% and 68.4%, respectively. The specificity of these tests was 96.6%, 98.3%, and 83.3%, respectively. When all 4 tests were assessed together, it was found that the sensitivity had risen to 94.7%. When the ELISA-kit was assessed with the IHA-kit and IHA-TA together, it was found that the sensitivity was 89.5% and 84.2%, respectively. Likewise, the combination of the ELISA-lab and IHA-kit or IHA-TA allowed us to achieve a sensitivity of 84.2% in cases of pulmonary echinococcosis. In conclusion, the diagnosis would be imminent if least 2 tests were applied together.

Improvement of indirect enzyme-linked immunosorbent assay for detection of Japanese encephalitis virus antibodies in swine sera

  • Yang, Dong-Kun;Kim, Ha-Hyun;Jo, Hyun-Ye;Lee, Seung Heon;Jang, Sang-Ho;Lee, Sang-Oh;Choi, Sung-Suk;Cho, In-Soo
    • 대한수의학회지
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    • 제57권1호
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    • pp.31-36
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    • 2017
  • Japanese encephalitis (JE) is an important zoonosis caused by the mosquito-transmitted JE virus (JEV), which is a causative agent of reproductive failure in pregnant sows. Detection of JEV antibodies in swine is performed by hemagglutination inhibition (HI), virus neutralization (VN), and the plaque reduction neutralization test (PRNT). The most stringent PRNT is the 90% endpoint PRNT ($PRNT_{90}$). These conventional assays are difficult to carry out in diagnostic laboratories with insufficient instruments or cell culture systems. An alternative assay that is easily conducted and time efficient is required. In this study, we improved the indirect enzyme-linked immunosorbent assay (I-ELISA) with clarified antigen for the detection of JEV antibodies. The I-ELISA results obtained from 175 swine serum samples were compared with HI, VN, and $PRNT_{90}$ results. The sensitivity of I-ELISA was 91.8%, 95.0%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. The specificity of I-ELISA was 92.2%, 94.7%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. Moreover, the I-ELISA results were significantly correlated with the HI (r = 0.93), VN (r = 0.95), and $PRNT_{90}$ (r = 0.92) results. These results suggest that the improved I-ELISA is useful for serosurveillance of JEV in swine.

전북지역 양돈장에서의 돼지 부종병 항체 및 톡신 양성률 조사 (Prevalence of antibody and toxin against edema disease from pig farms in Jeonbuk province)

  • 조선영;유정희;유영주;이한준;허진
    • 한국동물위생학회지
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    • 제46권4호
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    • pp.325-334
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    • 2023
  • Edema disease (ED) in pigs is enterotoxemia caused by Shiga toxin type 2e (Stx2e)-producing Escherichia coli (STEC) and frequently occurs in young piglets. Therefore, ED causes enormous economic losses in pig farms. In this study, a modified Stx2e (mStx2e) antigen was expressed and purified using commercial E. coli expression system. Monoclonal antibody was serviced by Ynto Ab Inc., using Phage Display Technique. Anti-Stx2e antibodies in piglets were measured by indirect ELISA using mStx2e antigens. Naive Stx2e in piglets were detected by Sandwich ELISA using Stx2e-monoclonal antibodies and commercial Stx2e-polyclonal antibodies. Among 3,480 piglets, anti-Stx2e antibodies were observed in 2,573 piglets. The 49.4% among 830 piglet serum samples possessed 0.625 ㎍/mL or more of Stx2e proteins. The 18.3% of 830 sera had 0.313 ㎍/mL of Stx2e proteins. The 32.3% of 830 samples held 0.156 ㎍/mL or less of Stx2e proteins. These results show that indirect ELISA using mStx2e antigen and Sandwich ELISA using Stx2e-monoclonal and polyclonal antibodies can be useful to detect ED in piglets.