• Parasites, Hosts and Diseases
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• 제21권2호
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• pp.270-280
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• 1983

임상적으로 폐흡충증이 의심되나 충란검출이 안되었던 155예(3예는 충란발견)의 피내반응 양성례에 대하여 Agar-gel Diffusion (AGD)반응, Counterimmunoelectro-phoresis(CIEP)반응 및 Enzyme-Linked Immunosorbent Assay(ELISA)를 실시하여 이들사이의 상관관계를 검토하였다. AGD 반응과 CIEP 반응은 폐흡충친항원원액(단백농도 7.56mg1m1)과 혈청원연을 사용하였고 ELISA반응에서는 상기 조항원의 40,000배 희석항원을, 그리고 혈청은 100배 및 200배 희석혈청을 사용하였다. 1. AGD 반응과 CIEP 사이의 양성반응의 일치률은 98%이었고 융성반응의 일치률은 100%이었다. 침강대는 AGD 반응에서 1∼3개가 흔히 관찰되었고 CIEP 반응에서는 1∼7개가 관찰되었으며 염색강도도 후자에서 일부의 피검혈청에서 강하게 나타났다. 2. AGD 반응과 ELISA 반응은 사성반응에서는 100배 희석혈청에서 96%, 그리고 200배 희석혈청에서 94%의 일치률을 나타내었고, 음성반응에서는 100석 희석혈청 및 200배 회석혈청에서 각각 97%와 99%의 일치율을 나타내었다. 3. CIEP와 ELISA 반응은 양성반응에서 100배 희석혈청에서는 98%, 200배 희석혈청에서는 96%의 일치률을 나타내었고 융성반응에서는 100배 희석혈청에서 97%, 200배 희석혈청에서 99%의 일치률을 나타내었다. 4. 대조혈청인 간흡충감염 혈청, 아메바감염 혈청 및 Toxoplaima형광항체반응 양성혈청에 대한 AGD 반응, CIEP 및 ELISA 반응은 전례에서 음성이었다. 이상의 결과로 볼 때 반응에 사용된 항원-항혈청의 농도로 보아서는 ELISA 반응의 감수성이 월등히 높았다. 같은 농도의 항원과 혈청을 사용한 AGD와 CIEP 반응에서는 CIEP에서 침강반응의 염색강도가 보다 강하게 나타난 것으로 보아 세가지 반응의 감수성의 높은 순위는 ELISA, CIEP, AGD라고 보여진다. 그러나 본 실험에서 AGD와 CIEP반응에서 ELISA보다 높은 농도의 항원과 항혈청을 사용하였을 때 감수성의 차이는 거의 볼 수 없는 유사한 상관관계의 반응을 나타내었다. 즉 반응방법에 따라 적절한 농도의 항원과 항혈청을 사용한다면 감수성의 차이는 없다고 본다. 세가지 반응의 유효성 으로 볼 때 가장 수기가 간편한 AGD 반응은 조항원을 사용할 때 용이하게 사용할 수 있는 방법이라 생각되며 CIEP는 AGD보다 전기영동장치가 필요한 번거로움이 있으나 보다 조기에 반응 결과를 알 수 있다. 가장 감수성이 예민한 ELISA반응은 항원을 정제하여 소동의 항원으로 다수의 집권집사를 할 수 있는 가장 바람직한 방법이라 생각된다. (본 실험을 수행함에 있어 ELISA 반응을 검사해 주신 연세의대 기생충학교실의 장재경 선생님께 감사드립니다)

• Parasites, Hosts and Diseases
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• 제28권4호
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• pp.207-212
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• 1990

이락촌폐흡충(Paragonimusiloktsuenensis) 감염 흰쥐 혈청의 항체 생산을 효소면역 반응 (enzyme-linked immunosorbent assay: ELISA)으로 관찰하였다. 이락촌폐흡충의 피낭유충을 흰쥐 에 감염시킨 후 격주로 12주까지 채질한 혈청과 이락촌폐흡충 성충 추출 조항원의 반응, 면역교차 반응의 유무를 알기 위해 동혈청과 폐흡충(P. westermani) 조항원(PWA) 및 간흡충(Clonorchis sinensis) 조항원(CSA)의 반응을 ELISA의 OD값(optical density value)으로 측정하였다. 감염강도 별 혈청군(GI, GII, GIII, 및 GIV)은 회수된 충체수로 구분하였다. 이락촌폐흡충 감염 횐쥐 혈청은 동항원(PIA)과의 반응에서 감염 후 4주에 전 감염 개체의 평균 OD값이 대조 혈청과 유의한 차이를 나타내어 항체 양성 반응이 인정되었고, 조사된 12주까지 점차 OD간의 증가로 항체가의 증가를 나타내었다. 감염강도별로 구분한 군별 평균 OD값의 수치는 대체로 감염강도와 비례되었으나 감염 6주 이후 군별 OD값의 유의성은 없었고, 개체별 OD값은 보유충체 수에 비례하지 않는 개체들이 많았다. 이락촌폐흡충 감염 횐쥐 힐청은 폐흡충(p. westermani) 조항원(PWA) 및 간흡충(C. sinensis)조항원(CSA)과의 반응(ELISA)에서도 상당한 면역교차 반응을 나타내었다.

• 대한수의학회지
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• 제39권1호
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• pp.159-168
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• 1999

Swine whipworm(Trichuris suis) is cosmopolitan nematode which can cause serious pathology in immature stage(larva2~larva5) of infected pigs, such as anorexia, diarrhea, anemia, and death in heavy infections. In this larval stages, it is very difficult to diagnose the infection of whipworm and to differentiate from other common swine gastrointestinal disorders such as 21 day scours which are associated with TGE virus, rota virus, coccidium, and the stress of weaning. In this experiment, the isolated antigens of Trichuris spp. were carried out to examine the structure and specificity of antigens and to select the reasonable antigens which would be used in serological diagnosis by electrophoresis, Western blotting, ELISA. The results of this experiment were as follows; 1. The common fractions of each Trichuris suis antigen were identified 28,32,45, 80kDa by SDS-PAGE with silver stain and four major fractions could be detected in positive swine sera by Western blot analysis. 2. The OD(optical density) values of somatic and excretory-secretory antigens which were reacted against positive(negative) sera from pigs infected with Trichuris suis by ELISA reader were; 1) OD values($mean{\pm}SD$) of adult somatic antigen against positive(negative) sera were $0.30{\pm}0.12(0.09{\pm}0.006)$ and third-stage larva of somatic antigen were $0.28{\pm}0.038(0.10{\pm}0.005)$. And OD values of excretory-secretory antigens of adult and third-stage larva were $0.24{\pm}0.031(0.11{\pm}0.005)$ and $0.08{\pm}0.013(0.10{\pm}0.003)$, respectively. 2) OD values of adult somatic, larval somatic antigen and adult excretory-secretory antigen response to positive sera were significantly (p<0.01) associated with negative swine sera. And the Cut-off OD values(minimum positive value) were determined to be mean negative value plus 3 SD that would minimized the risk of false positives. 3. The OD values of somatic antigens of T suis and T vulpis against swine positive(negative) sera were $0.30{\pm}0.120(0.09{\pm}0.006)$ and $0.25{\pm}0.141(0.09{\pm}0.003)$. These data mean that the somatic antigens of T suis and T vulpis were able to diagnose T vulpis infection in dogs as well as T suis infection in pigs. These results suggest that somatic antigen of third-stage larva and excretory-secretory antigen of adult T suis could be used the diagnostic antigen by serological test(ELISA) in immature Trichuris spp. infection.

• Parasites, Hosts and Diseases
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• 제36권1호
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• pp.37-46
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• 1998

인체 간흡충증의 현증감염 시기에 생산된 특이 IgG 항체와 반응하는 간촘충 특이항원을 규명하였다. 간흡충 성충의 조항원 및 서로 다른 조건에서 회수한 4가지 분비배설항원 (CsE)의 항원성을 면역이적법 소견으로 비교하였다 CsE2의 30. 7 kDa 항원이 간흡충 환자혈청과 강한 면역반응을 보였다. 이 항원들은 프라지콴텔 완치 후 6개월 혈청과는 반응하지 않았으나 투약 후 간흡충 충란이 검출된 환자의 치료 후 혈청과는 반응하여 현증감염의 면역반응과 관련된다고 판단하였다. 그러나. 30 Ka항원은 폐흡충 요코가와흉충 감염혈청과 교차반응하였고. 7 kDa 항원은 반응하지 않았다. 또. 30 kDa 항원은 일부 간흡충 과거 감염자 혈청과 반응하였으나 7 kDa 항원은 반응하지 않아 7 님)a 항원의 특이도가 높았다 간흡충 유행지역에서 7 kDa 항원의 민감도를 파악한 바 간흡충 현증감염자 25명 중 23명 (92%) 간홉충 요코가와흡충 중복감염자 11명 중 10명 (91%)과 반응하였다. 반면 간흡충 과거감염자 15명 중 6명 (40%), 요코가와흡충 감염자 7명 중 3명 (43%)과 반응하였다. 따라서. CsE2의 7 kDa 항원은 인체 간흡충 현증감염의 표식자가 되는 간흡충 특이 항원으로 판단된다.

• 대한수의학회지
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• 제38권4호
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• pp.859-866
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• 1998

This study was carried out to investigate the prevalence of Neopsora (N) caninum infection in Korean dairy herds. To determine the prevalence of antibodies to N caninum in Korean dairy cattle, a total of 1,688 sera including 895 sera taken from 30 herds having recent high abortion rate and 793 sera selected randomly from 168 herds with no history of recent abortion problem, respectively, collected nationwide during a designated period were analyzed by indirect fluorescent antibody (IFA) test. Mean nationwide seropositive rate of the sera tested in herds and individual cattle tested were 53.5% and 35.6%, respectively. However mean seropositive rate of the samples from herds having abortion problem was approximately two and half times higher than those in herds with no recent abortion history. Regional seropositive rates of the samples from the herds with abortion problem were 48.6%, 51.6%, 44.4% and 71.4% at Kyunggi, Kangwon, Kyungbuk and Jeonnam province, respetively. Regional seropositive rates of the samples from the herds with no recent abortion problem were 35.6%, 18.3%, 16.5%, 37.5%, 19.4%, 33.3%, 32.1%, 3.8% and 0.0% at Kyunggi, Kangwon, Chungbuk, Chungnam, Kyungbuk, Kyungnam, Jeonbuk, Jeonnam and Jeju province, respectively. The results of this study suggested that N caninum infection was widespread and considerably associated with bovine abortion in Korea.

• Parasites, Hosts and Diseases
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• 제47권3호
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• pp.219-225
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• 2009

The seroprevalence of cryptosporidiosis was examined using patients' sera collected from hospitals located in 4 different areas of the Republic of Korea. ELISA was used to measure antibody titers against Cryptosporidium parvum antigens from a total of 2,394 serum samples, which were collected randomly from patients in local hospitals; 1)Chungbuk National University Hospital, 2) Konkuk University Hospital, 3) local hospitals in Chuncheon, Gangwon-do (province), 4) Jeonnam National University Hospital, from 2002 through 2003. Of the2,394 samples assayed, 34%, 26%, and 56% were positive for C. parvum-specific IgG, IgM, and IgA antibodies, respectively, Positive IgG titers were most common in sera from Jeonnam National University Hospital, Gwangju, Jeollanam-do, and positive IgM titers were most common in sera from Chungbuk National University Hospital, Cheongju, Chuncheonqbuk-do. The seropositivity was positively correlated with age for both the IgG and IgA antibodies but was negatively correlated with age for the IgM antibodies. Western blotting revealed that 92%, 83%, and 77% of sera positive for IgG, IgM, and IgA ELISA reacted with 27-kDa antigens, respectively. These results suggested that infection with Cryptosporidium in hospital patients occurs more commonly than previously reported in the Republic of Korea.

• Biomolecules & Therapeutics
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• 제1권2호
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• pp.236-243
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• 1993

Immunologic potential of DA-125, a new anthracycline antitumor antibiotic, was investigated using guinea pigs and mice. In antigenicity experiments, guinea pigs were sensitized subcutaneously with DA-125 or DA-125 incorporated in complete Freund's adjuvant (CFA) once a week for three weeks. No systemic anaphylaxis was induced by intravenous injection of DA-125 or DA-125 incubated with guinea pig serum after 3 weeks from the last sensitization. None of sera of these animals showed any passive cutaneous anaphylactic reaction (PCA) when DA-125 or DA-125 incubated with guinea pig serum was used as a challenging antigen in homologous PCA experiment. On the other hand the treatment of guinea pigs with ovalbumin Incorporated in CFA induced systemic anaphylactic reaction when challenged by intravenous injection of 5 mg/body of ovalbumin. Immunodiffusion test revealed no precipitating antibodies as detected in guinea pigs sensitized with DA-125. In 24-hour heterologous PCA reaction with sera of C57BL/6 mice immunized with DA-125 or DA-125 mixed with aluminum hydroxide gel (Alum), None of sera showed positive reaction when DA-125 or DA-125 incubated with rat serum was used as a challenging antigen. Sera of animals immunized with a mixture of ovalbumin and alum showed positive PCA reaction when 5 mg/body of ovalbumin was injected as a challenging antigen. In lymphocyte proliferation tests, spleen lymphocyte proliferation to PHA and LPS was similarly impaired by 12 mg/kg of DXR or 36 mg/kg of DA-125, and the immunodepressive activity of DA-125 showed a dose-dependent manner. From these results, it could be concluded that immunosupression of DA-125 would be comparable to that of DXR and that DA-125 would not induce systemic allergic reaction in its clinical use.

• Parasites, Hosts and Diseases
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• 제38권2호
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• pp.75-84
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• 2000

Antibody responses in serum and cerebrospinal fluid (CSF) samples from patients with active and chronic paragonimiasis and in sera from patients on whom follow-up studies were done after praziquantel treatment were analyzed using antigens of Paragonimus westermani prepared from eggs, metacercariae, juveniles of 4-and 7-week old, adult worms and recombinant protein of 28 kDa cruzipain-like cysteine protease (rPw28CCP). The patient sera/CSFs of active and chronic paragonimiasis revealed strong antibody reactions against the crude extracts of 4-and 7-week old juveniles as well as against those from egg and adult. rPw28CCP also showed specific reaction to the sera with active paragonimiasis. After the treatment, levels of specific antibodies in the sera gradually decreased to negative range in most patients. In some cases with persisting high antibody levels, however, the reactions at 27 kDa egg Protein were sustained throughout the observation period of 34 months. The reactions at 35 and 32 kDa in adult extract and rPw28CCP disappeared rapidly after the treatment. Persistent antibody reactions even after successful treatment are provoked by continuous antigenic challenge from eggs which were not resolved by treatment.

• Parasites, Hosts and Diseases
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• 제51권6호
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• pp.763-766
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• 2013

A synthetic peptide was prepared based on the antigenic region of Paragonimus westermani pre-procathepsin L, and its applicability for immunodiagnosis for human paragonimiasis (due to Paragonimus heterotremus) was tested using an ELISA to detect IgG4 antibodies in the sera of patients. Sera from other helminthiases, tuberculosis, and healthy volunteers were used as the references. This peptide-based assay system gave sensitivity, specificity, accuracy, and positive and negative predictive values of 100%, 94.6%, 96.2%, 100%, and 88.9%, respectively. Cross reactivity was frequently seen against the sera of fascioliasis (75%) and hookworm infections (50%). Since differential diagnosis between paragonimiasis and fascioliasis can be easily done by clinical presentation and fascioliasis serology, this cross reaction is not a serious problem. Sera from patients with other parasitoses (0-25%) rarely responded to this synthetic antigen. This synthetic peptide antigen seems to be useful for development of a standardized diagnostic system for paragonimiasis.

• Parasites, Hosts and Diseases
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• 제51권4호
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• pp.485-488
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• 2013

The performance values of available techniques used in serodiagnosis of toxoplasmosis are satisfactory but they raise problems of equivocal and discordant results for very low IgG titers. Recently marketed, LDBio-Toxo II IgG Western blot (IB) showed an excellent correlation with the dye test. We estimated the proportion of equivocal and discordant results between the enzyme immunoassay Platelia Toxo IgG (EIA-IgG) and fluorescent antibody test (FAT) and assessed the usefulness of the IB as a confirmatory test. Out of 2,136 sera collected from pregnant women, 1,644 (77.0%) tested unequivocally positive and 407 (19.0%) were negative in both EIA-IgG and FAT. The remaining 85 (4%) sera showed equivocal or discordant results. Among them, 73 (85.9%) were positive and 12 (14.1%) were negative in IB. Forty-one (89.1%) equivocal sera in EIA-IgG and 46 (86.8%) equivocal sera in FAT were positive in IB. Reducing the cut-off values of both screening techniques improved significantly their sensitivity in detecting very low IgG titers at the expense of their specificity. In conclusion, equivocal results in routine-used techniques and their discordance in determination of the immune status in pregnancy women were not uncommon. IB test appeard to be highly useful in these situations as a confirmatory technique.