• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7345-7349
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• 2013

Background: Because of the high prevalence of hepatocellular carcinoma (HCC) in Egypt, new markers with better diagnostic performance than alpha-feto protein (AFP) are needed to help in early diagnosis. The aim of this work was to compare the clinical utility of both serum and mRNA glypican3 (GPC3) as probable diagnostic markers for HCC among Egyptian patients. Materials and Methods: A total of 60 subjects, including 40 with HCC, 10 with cirrhosis and 10 normal controls were analyzed for serum GPC3 (sGPC3) by ELISA. GPC-3 mRNA from circulating peripheral blood mononuclear cells was amplified by RT-PCR. Both markers were compared to some prognostic factors of HCC, and sensitivity of both techniques was compared. Results: Serum glypican-3 and AFP were significantly higher in the HCC group compared to cirrhotic and normal controls (p<0.001). Sensitivity and specificity were (95% each) for sGlypican-3, (82.5% and 85%) for AFP, and (100% and 90%) for Glypican3 mRNA, and (80% and 95%) for double combination between sGPC3 and AFP respectively. Conclusion: Both serum GPC-3 and GPC-3mRNA are promising diagnostic markers for early detection of HCC in Egyptian patients. RT- PCR proved to be more sensitive (100%) than ELISA (95%) in detecting glypican3.

• 대한수의학회지
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• 제37권3호
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• pp.583-593
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• 1997

Indirect fluorescent antibody test(IFAT) and enzyme-linked imuunosorbent assay (IgG-ELISA) as serological diagnostic tools were conducted to evaluate the usefulness for diagnosis of canine babesiosis infected with Babesia gibsoni in domestic various dog breeds, american pit bullterrier, military shepherd, and mongrel dogs. The results obtained from this study were abstracted as follows. The nonionic detergent Triton X-100 and absorbent bio-bead $SM_2$ were useful reagents for the preparation of pure merozoite antigen of B gibsoni to be used in ELISA. The optimum reaction in ELISA was shown when the protein concentration of ELISA antigen was measured as 625ng/ml and the conjugate concentration was diluted into 1/6000 fold. The average OD value of ELISA in sera determined with negative responses in IFAT was measured as $0.255{\pm}0.051$(490nm) and the cut - off value of OD was determined as 0.399(490nm). The serum antibodies in both of IFAT and ELISA were detected on one week after artificially infected with B gibsoni and these high antibody titers, 512X in IFAT and 1024X in ELISA, were long lasted until 15 weeks after infection. The reproducibility of reaction and stability of the antigen absorbed microtitration polystyrene plate preserved in $4^{\circ}C$ refrigerator and $-20^{\circ}C$ freezer, respectively could be lasted until 135 days after storage. The positive rates in IFAT by dog breeds were shown 8.1%(60/744 heads) in mongrel dogs, 81.3%(78/96 heads) in american pit bullterrier and 15.6%(15/96 heads) in military shepherd, while the positive rate in ELISA shown 17.6%(131/744 heads) in mongrel dogs, 83.3%(80/96 heads) in american pit bullterrier and 36.5%(35/96 heads) in military shepherd, respiectively. In the total of 936 heads surveyed with IFAT and ELISA the positive rates in IFAT and ELISA were 16.4%(153/936 heads) and 26.3%(246/936 heads), respectivily. Agreement of reactions between IFAT and ELISA was shown 82.4% in 936 dog sera. The specificity and sensitivity of ELISA reaction were 83.5% and 76.5%, respectively. From the conclusion obtained in this study it was evaluated that IFAT and ELISA were useful as highly specific, sensitive and stable serelogical tools for the diagnosis of canine babesiosis in Korea.

• Parasites, Hosts and Diseases
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• 제52권2호
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• pp.137-142
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• 2014

Serologic tests are widely accepted for diagnosing Toxoplasma gondii but purification and standardization of antigen needs to be improved. Recently, surface tachyzoite and bradyzoite antigens have become more attractive for this purpose. In this study, diagnostic usefulness of 3 recombinant antigens (SAG1, SAG2, and SAG3) were evaluated, and their efficacy was compared with the available commercial ELISA. The recombinant plasmids were transformed to JM109 strain of Escherichia coli, and the recombinants were expressed and purified. Recombinant SAG1, SAG2, and SAG3 antigens were evaluated using different groups of sera in an ELISA system, and the results were compared to those of a commercial IgG and IgM ELISA kit. The sensitivity and specificity of recombinant surface antigens for detection of anti-Toxoplasma IgG in comparison with commercially available ELISA were as follows: SAG1 (93.6% and 92.9%), SAG2 (100.0% and 89.4%), and SAG3 (95.4% and 91.2%), respectively. A high degree of agreement (96.9%) was observed between recombinant SAG2 and commercial ELISA in terms of detecting IgG anti-Toxoplasma antibodies. P22 had the best performance in detecting anti-Toxoplasma IgM in comparison with the other 2 recombinant antigens. Recombinant SAG1, SAG2, and SAG3 could all be used for diagnosis of IgG-specific antibodies against T. gondii.

• Parasites, Hosts and Diseases
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• 제53권6호
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• pp.699-703
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• 2015

Cystic echinococcosis (CE) is one of the most widespread zoonotic helminthiases, which can last an asymptomatic infection for several years. The purpose of this study was to demonstrate serum antibody prevalence of CE among asymptomatic people in Uzbekistan using ELISA. A total of 2,547 serum samples were collected, 66 from confirmed CE patients and 2,481 of patients with other diseases than CE at a hospital in Tashkent, Uzbekistan. The serum samples were screened for CE specific IgG antibodies by ELISA using cystic fluid antigen obtained from sheep. The serum antibody positive rate was 89.4% (59/66) in CE and 3.6% (89/2,481) in other disease patients. The present ELISA recognized 89.4% sensitivity and 96.4% specificity. The ELISA absorbance of positive samples was distributed 0.271-0.971 for CE and 0.273-0.887 for other disease patients. The other disease patients with high absorbance over 0.3 were 50 (2.0%) who were presumed to be active CE patients. The patients in their 40s showed the highest positive rate of 5.2% (P=0.181), and women were 4.4% while men were 3.1% positive (P=0.136). The data confirmed that there are many asymptomatic patients of CE in Tashkent. It is indicated that CE is an endemic disease of public health importance in Uzbekistan.

• Parasites, Hosts and Diseases
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• 제38권4호
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• pp.251-256
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• 2000

There have been some reports about the prevalence of anti-Toxoplasma gondii antibody among Koreans, and most of all data were taken from patients visiting hospitals. However, the epidemiological data of the community-based study in Korea are rare. This study was performed to evaluate the seroprevalence of toxoplasmosis among the inhabitants of the rural area Okcheon-gun, Korea. A total of 1,109 serum samples (499 males, 610 females) were examined for the IgG antibodies by ELISA. To set up the cut-off point for ELISA, we used a commercial latex agglutination (LA) kit. The sensitivity and specificity of ELISA against LA test were 89.5%, and 98.6% respectively. Among 1,109 sera, 6.9% showed seropositivity by ELISA. The positive rates of males and females were 6.0% and 7.2%, respectively. However, there were no significant differences between sexes. Comparing the age groups, the highest seropositive rate showed in the seventies or higher, and their rates had a tendency to increase with age (0.05 < p < 0.3). These results revealed that the seroprevalence of toxoplasmosis in rural inhabitants is similar to previous reports in Korea; however we need further investigation to clarify the prevalence of toxoplasmosis in the general population.

• Parasites, Hosts and Diseases
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• 제29권3호
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• pp.293-310
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• 1991

우리 나라의 중요한 인체 기생충인 간흡충(Cloncrchis sinensis)을 연구함에 있어, 단세포를 항체제조법을 이용하여 보았다. 간흡충의 성충 조(조)항원으로 면역한 마우스의 비장 림프구와 형질세포종(plasmacytoma) 세포를 융합하여 간흡충의 항원에 대한 단세포를 항체를 분비하는 융합 세포를 만들어, 간흡충 항원에 대한 특이 단세포군 항체를 얻은 후, 이의 특성 및 반응하는 항원의 특성을 분석하고, 아울러 ELISA 억제 검사법을 이용하여 면역 진단법 상 특이도의 개선을 모색하였다 그 결과, 간흡충 항원에 대한 항체를 분비하는 총 29개의 융합세포군을 확인하였는데, 이 중 8개는 다른 기생 충 항원에 대해 교차 반응을 나타내지 않는 높은 특이성을 지니고 있었다. 클로닝 후 선택된 6종류의 단세포군 항체는 Ig Gl이 넷이었고, 나머지는 Is G2b 및 Is A로 나타났다. 위와 같이 선택된 단세포를 항체 중 4개만이 자연 감염 시에도 표현되는 항원 결정기에 대하여 생성된 것으로 판단되었다. 효소 면역 전기영동 블로팅으로 분자량 l0KD, 34 KD 항원은 각각 CsHyb 0714-20 및 CsHyb 0605-10단세포군 항체와 항원항체 반응을 나타냄을 확인하였다. 간접형광항체법을 이용하여 카 항원 결정기의 충체 내 위치를 관찰한 결과, CsHyb 0714-20 단세포를 항체에 대한 항원은 충체의 표면 및 실질 대부분에 분포하였으며, CsHyb 0605-10 및 CsHyb 0714-25 단세포군 항체에 대한 항원은 충체의 실질 및 장관의 상피세포에 분포하고 있었다. 한편 CsHyb 0605-23 단세포군 항체에 대한 항원은 주로 자궁내 충란 주위에 많이 분포하고 있었다. 단세포군 항체와 반응하는 Sephadex G200 겔 여과 항원 분획을 검색한 결과, 검색한 항원 결정기는 모두 전반적으로 빨리 젤 여과를 통하여 나온 분회에 속하여 있는 것을 알 수 있었다. 한편 이 단세포군 항체를 인체 간흡충증의 진단에 응용하여 그 특이도를 개선하고자 하였다. 통상적인 방법의 ELISA로 시행한 항체가로는 간흡충 감염자의 75U가 양성 병위에 들었으며, 정상 대조군의 7.l%, 폐흡충 감염 자의 37.5%가 위 양성 반응을 보였다. 반면 CsHyb 0605-23 단세포군 항체를 함께 사용하여 ELISA억제 검사를 시행한 결과는 간흡충 감염자의 77.1%가 양성으로 판정되었으며, 정상 대조군 및 폐흡충 감염자에서는 양성 반 응을 찾아 볼 수 없어서 100%의 특이도를 나타내었다. 따라서 이러한 단세포군 항체를 이용한 ELISA 억제검사 는 통상적인 ELISA 검사에 비하여 같은 정도의 민감도를 유지하면서도 매우 높은 특이도를 가지는 것으로 판단되었다.

• 한국식품위생안전성학회지
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• 제33권3호
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• pp.185-192
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• 2018

본 연구에서는 수산가공품 중 고등어 어육을 신속하게 검출하기 위하여 고등어 어육 중 열 안정-수용성 단백질에 특이한 단클론성 항체(3A5-2)를 이용하여 indirect ELISA 법을 개발하였다. Indirect ELISA을 개발하기에 앞서 먼저 시료 전처리는 이전 연구의 결과를 바탕으로 진행되었다. 이전 연구에서는 3A5-2 항체가 37 kDa 부근의 열 안정-수용성 단백질과 반응하였고, 0.05 M carbonate buffer로 추출하였을 때 흡광도가 가장 두드러지게 증가한 것을 확인하였다. 따라서 indirect ELISA의 시료 전처리는 0.05 M carbonate buffer를 이용한 열처리 추출법으로 추출한 후 0.05 M PBS로 희석하는 것으로 확립하였고, indirect ELISA 법을 최적화하였다. 개발된 indirect ELISA법에 실험실에서 임의로 열처리한 고등어 샘플을 적용한 결과 indirect ELISA법은 0.001% (0% 흡광도 표준편차 양의 값: $0.0003{\times}3$)의 검출한계를 확인하였으며, 꽁치 중 고등어는 0.002%(0% 흡광도 표준편차 양의 값: $0.0006{\times}3$)까지 검출이 가능하였다. 또한 조리($100^{\circ}C$, 30분)와 멸균($121^{\circ}C$, 30분) 처리된 고등어의 검출이 가능한 것으로 확인되었고, 멸균된 제품에서도 고등어에 특이적으로 반응하여 고등어의 혼입여부 판별도 가능한 것으로 판단되었다. 시판되는 고등어 가공품에 대해서는 양성결과를 나타내었고 다른 수산물에서는 음성으로 판단되어 개발된 분석법은 가공품에 혼입될 수 있는 알레르겐인 고등어를 보다 신속하고 민감하게 분석할 수 있고, 점차 가격이 증가하고 있는 고등어의 혼입여부를 확인할 수 있는 분석 도구로서 활용이 가능할 것으로 판단된다.

• Parasites, Hosts and Diseases
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• 제56권1호
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• pp.61-70
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• 2018

We developed a Rapid Diagnostic Test (RDT) kit for detecting IgG/IgM antibodies against Zika virus (ZIKV) using monoclonal antibodies to the envelope (E) and non-structural protein 1 (NS1) of ZIKV. These proteins were produced using baculovirus expression vector with Sf9 cells. Monoclonal antibodies J2G7 to NS1 and J5E1 to E protein were selected and conjugated with colloidal gold to produce the Zika IgG/IgM RDT kit (Zika RDT). Comparisons with ELISA, plaque reduction neutralization test (PRNT), and PCR were done to investigate the analytical sensitivity of Zika RDT, which resulted in 100% identical results. Sensitivity and specificity of Zika RDT in a field test was determined using positive and negative samples from Brazil and Korea. The diagnostic accuracy of Zika RDT was fairly high; sensitivity and specificity for IgG was 99.0 and 99.3%, respectively, while for IgM it was 96.7 and 98.7%, respectively. Cross reaction with dengue virus was evaluated using anti-Dengue Mixed Titer Performance Panel (PVD201), in which the Zika RDT showed cross-reactions with DENV in 16.7% and 5.6% in IgG and IgM, respectively. Cross reactions were not observed with West Nile, yellow fever, and hepatitis C virus infected sera. Zika RDT kit is very simple to use, rapid to assay, and very sensitive, and highly specific. Therefore, it would serve as a choice of method for point-of-care diagnosis and large scale surveys of ZIKV infection under clinical or field conditions worldwide in endemic areas.

• 대한미생물학회지
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• 제21권2호
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• pp.181-189
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• 1986

To apply ELISA to serodiagnosis of leptospirosis with killed whole cells from Leptospira interrogans serovars mwogolo (Mwogolo), copenhageni (M-20), WH-20, autumnalis (Akiyami A), cynopteri (3522 C), australis (Bacillico) and Leptospira biflexa serovar patoc (patoc 1), sensitivity and specificity was evaluated. The reactivity of IgM and IgG antibody in the sera from patients with leptospirosis, hemorrhagic fever with renal syndrome and other febrile disease and normal healthy control to the killed whole cells was analysed. The results were summarized as follows. 1. The reactivity (absorbance at 492mn) of IgM and IgG to L. mwogolo antigen in the sera of pattients with leptospirosis were $1.414{\pm}0.370$, $1.242{\pm}9.554$ respectively: hemorrhagic fever with renal syndrome, $0.329{\pm}0.131$, $0.239{\pm}0.126$; other febrile disease, $0.196{\pm}0.071$, $0355{\pm}0.141$; normal healthy control, $0.136{\pm}0.016$, $0.208{\pm}0.077$. 2. The reactivity of IgM and IgG to L. copenhageni, WH-20, L. autumnalis, L. cynopteri and L. anstralis antigens were similar to that to L. mwogolo antigen, but that to L. biflexa antigen was not discriminated among above disease. 3. Correlation coefficient between the MAT titer and ELISA OD (IgM) to the above antigens was in the range of 0.071-0.518. 4. As absorbance above 0.60 was determined positive for the diagnosis of leptospirosis, the sensitivity and specificity of IgG was 25-89% and 91-96% respectively. And those of IgM was 98-100% and 89-100% except L. biflexa (29%) respectively.

• Parasites, Hosts and Diseases
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• 제22권2호
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• pp.223-228
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• 1984

외과적으로 충체를 적출하여 진단한 스파르가눔증 환자 7명으로부터 혈청을 수집하고 혈청내 스파르가눔 특이 IgG 항체가를 호소면역 측정 법으로 측정하였다. 스파르가눔 항원은 유혈목이에 자쳔 감염된 충체를 갈아 생리 식염수로 추출한 것으로 단백질 함량은 Smgyxnl이었다. 효소면역측정법은 McLaren 등(1978)의 방법에 따라 실시하였다. 스파르가눔중 환자 혈청이 폐흡충 항원 또는 유구낭미충 항원에 교차반응이 있는지 여부도 측정하였다. 또한 스파르가눔 항원에 대해서 정상인, 케흡충중 환자, 간홉충 감염자, 유구낭미충증 환자와 무구조충 감염자 등 모두 71명에 대해서도 교차반응을 나타내는지 를 관찰하고자 효소면역 측정 법을 실시하였다. 그 결과를 요약하면 다음과 같다. 1. 스파르가늠증 환자 7명중 석회화한 스파르가눔에 감염되어 있었고 수술후 1개월에 혈청을 수집한 1례를 제외하면 모두 홉광도 0.3 이상을 나타내었다. 양성판정기준을 흠광도 0.25로 하였을 때 효소면역 측정 법의 민감도(sensitivity)는 85.7이 었다. 2. 기타 기생충 감염자와 정상인 71명중에서 스파르가눔 항원에 대한 특이항체 양성자는 무구조충감염자 3명으로 특이도(specificity)는 97.5%이었다. 3. 스파르가눔중 환자 혈청은 폐흡충 항원 및 유구낭미충 항원에 대해서 항체가는 낮은 값을 보이고 있어 교차 반응은 발견할 수 없었다. 4. 중추신경계를 침범한 스파르가눔중 환자 2례에서 뇌척수액을 희석하지 않은 상태로 효소면역 측정 법을 실시하였던 바 1례는 폐흡충 및 유구낭미충 항원에 대해 교차반응 없이 스파르가눔항원에 대해 높은 항체가를 보인데 비해 다른 1례에서는 스파르가눔 항원 및 유구낭미충 항원에 대해 비슷한 항체가를 나타내고 있었다. 이상의 결과에서 효소면역 측정 법을 이용하여 스파르가눔중을 혈청학적으로 진단하는 경우 매우 특이하고 민감한 결과를 얻을 수 있을 것으로 생각하였다. 스파르가눔중의 수술전 진단이 실제로 어려운 것이 현실이므로 혈청학적 잖단은 역학적 유해조사나 형태학적으로 감별이 어려운 병리소견을 보인 경우 등에서 보조적인 진단법으로 이용할 수 있을 것으로 생각한다.