• Title/Summary/Keyword: cross reactivity

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Development of a Rapid Automated Fluorescent Lateral Flow Immunoassay to Detect Hepatitis B Surface Antigen (HBsAg), Antibody to HBsAg, and Antibody to Hepatitis C

  • Ryu, Ji Hyeong;Kwon, Minsuk;Moon, Joung-Dae;Hwang, Min-Woong;Lee, Jeong-Min;Park, Ki-Hyun;Yun, So Jeong;Bae, Hyun Jin;Choi, Aeran;Lee, Hyeyoung;Jung, Bongsu;Jeong, Juhee;Han, Kyungja;Kim, Yonggoo;Oh, Eun-Jee
    • Annals of Laboratory Medicine
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    • v.38 no.6
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    • pp.578-584
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    • 2018
  • Background: Accurate, rapid, and cost-effective screening tests for hepatitis B virus (HBV) and hepatitis C virus (HCV) infection may be useful in laboratories that cannot afford automated chemiluminescent immunoassays (CLIAs). We evaluated the diagnostic performance of a novel rapid automated fluorescent lateral flow immunoassay (LFIA). Methods: A fluorescent LFIA using a small bench-top fluorescence reader, Automated Fluorescent Immunoassay System (AFIAS; Boditech Med Inc., Chuncheon, Korea), was developed for qualitative detection of hepatitis B surface antigen (HBsAg), antibody to HBsAg (anti-HBs), and antibody to HCV (anti-HCV) within 20 minutes. We compared the diagnostic performance of AFIAS with that of automated CLIAs-Elecsys (Roche Diagnostics GmbH, Penzberg, Germany) and ARCHITECT (Abbott Laboratories, Abbott Park, IL, USA)-using 20 seroconversion panels and 3,500 clinical serum samples. Results: Evaluation with the seroconversion panels demonstrated that AFIAS had adequate sensitivity for HBsAg and anti-HCV detection. From the clinical samples, AFIAS sensitivity and specificity were 99.8% and 99.3% for the HBsAg test, 100.0% and 100.0% for the anti-HBs test, and 98.8% and 99.1% for the anti-HCV test, respectively. Its agreement rates with the Elecsys HBsAg, anti-HBs, and anti-HCV detection assays were 99.4%, 100.0%, and 99.0%, respectively. AFIAS detected all samples with HBsAg genotypes A-F and H and anti-HCV genotypes 1, 1a, 1b, 2a, 2b, 4, and 6. Cross-reactivity with other infections was not observed. Conclusions: The AFIAS HBsAg, anti-HBs, and anti-HCV tests demonstrated diagnostic performance equivalent to current automated CLIAs. AFIAS could be used for a large-scale HBV or HCV screening in low-resource laboratories or low-to middle-income areas.

Study on the characteristics of transition metals for TSSG process of SiC single crystal (SiC 단결정의 TSSG 공정을 위한 전이금속 특성 연구)

  • Lee, Seung-June;Yoo, Yong-Jae;Jeong, Seong-Min;Bae, Si-Young;Lee, Won-Jae;Shin, Yun-Ji
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.32 no.2
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    • pp.55-60
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    • 2022
  • In this study, a heat treatment experiment was conducted to select a new melt composition that can easily control the unintentionally doped nitrogen (N-UID) without degrading the SiC single crystal quality during TSSG process. The experiment was carried out for about 2 hours at a temperature of 1900℃ under Ar atmosphere. The used melt composition is based on either Si-Ti 10 at% or Si-Cr 30 at%, and also Co or Sc transition metals, which are effective for carbon solubility, were added at 3 at%, respectively. After the experiment, the crucible was cross-sectionally cut, and evaluated the Si-C reaction layer on the crucible-melt interface. As a result, with Sc addition, Si-C reaction layers uniformly occurred with a Si-infiltrated layer (~550 ㎛) and a SiC interlayer (~23 ㎛). This result represented that the addition of Sc is an effective transition metal with high carbon solubility and can feed carbon sources into the melt homogeneously. In addition, Sc is well known to have low reactivity energy with nitrogen compared to other transition metals. Therefore, we expect that both growth rate and Nitrogen UID can be controlled by Si-Sc based melt in the TSSG process.

Development of a lateral flow dipstick test for the detection of 4 strains of Salmonella spp. in animal products and animal production environmental samples based on loop-mediated isothermal amplification

  • Wirawan Nuchchanart;Prapasiri Pikoolkhao;Chalermkiat Saengthongpinit
    • Animal Bioscience
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    • v.36 no.4
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    • pp.654-670
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    • 2023
  • Objective: This study aimed to develop loop-mediated isothermal amplification (LAMP) combined with lateral flow dipstick (LFD) and compare it with LAMP-AGE, polymerase chain reaction (PCR), and standard Salmonella culture as reference methods for detecting Salmonella contamination in animal products and animal production environmental samples. Methods: The SalInvA01 primer, derived from the InvA gene and designed as a new probe for LFD detection, was used in developing this study. Adjusting for optimal conditions by temperature, time, and reagent concentration includes evaluating the specificity and limit of detection. The sampling of 120 animal product samples and 350 animal production environmental samples was determined by LAMP-LFD, comparing LAMP-AGE, PCR, and the culture method. Results: Salmonella was amplified using optimal conditions for the LAMP reaction and a DNA probe for LFD at 63℃ for 60 minutes. The specificity test revealed no cross-reactivity with other microorganisms. The limit of detection of LAMP-LFD in pure culture was 3×102 CFU/mL (6 CFU/reaction) and 9.01 pg/μL in genomic DNA. The limit of detection of the LAMP-LFD using artificially inoculated in minced chicken samples with 5 hours of pre-enrichment was 3.4×104 CFU/mL (680 CFU/reaction). For 120 animal product samples, Salmonella was detected by the culture method, LAMP-LFD, LAMP-AGE, and PCR in 10/120 (8.3%). In three hundred fifty animal production environmental samples, Salmonella was detected in 91/350 (26%) by the culture method, equivalent to the detection rates of LAMP-LFD and LAMP-AGE, while PCR achieved 86/350 (24.6%). When comparing sensitivity, specificity, positive predictive value, and accuracy, LAMP-LFD showed the best results at 100%, 95.7%, 86.3%, and 96.6%, respectively. For Kappa index of LAMP-LFD, indicated nearly perfect agreement with culture method. Conclusion: The LAMP-LFD Salmonella detection, which used InvA gene, was highly specific, sensitive, and convenient for identifying Salmonella. Furthermore, this method could be used for Salmonella monitoring and primary screening in animal products and animal production environmental samples.

Analysis of Antibodies Cross-reactive with Pressate Extract Antigen from Mycobacterium tuberculosis and Other 3 Species Mycobacteria in Sera of Patients with Pulmonary Tuberculosis (결핵균 및 기타 3종 Mycobacteria의 파쇄추출항원과 교차반응하는 폐결핵환자의 항체분석)

  • Cho, Myung-Je;Hwang, Eung-Soo;Kook, Yoon-Hoh;Kim, Ik-Sang;Lee, Seoung-Hoon;Cha, Chang-Yong;Shim, Young-Soo;Han, Yong-Chol;Bae, Gill-Han;Kim, Sang-Jae
    • The Journal of the Korean Society for Microbiology
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    • v.20 no.1
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    • pp.79-89
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    • 1985
  • It is important to discriminate between tuberculosis and tuberculosis-like disease by Mycobacteria other than tuberculosis in the serodiagnosis of tuberculosis. But because common antigens share among Mycobacteria, their antigenicities to human are similar. Therefore degree of cross-reactivity of antibody in the sera of patients with tuberculosis between M. tuberculosis and Mycobacteria other than tuberculosis should be checked to increase the specificity in the serodiagnosis of tuberculosis. The activity levels of IgG antibody in the sera of 106 patients confirmed as active pulmonary tuberculosis and 30 normal healthy control person to the pressate extract antigen (TE, BE, AE, and FE antigen) from M. tuberculosis, M. bovis, M. avium, and M. fortuitum were measured by enzyme-linked immunosorbent assay and the crossreactivity of IgG antibody with mycobacterial species was analysed. The results were as follows; 1. The activity level(O.D. at 492nm) of IgG to TE antigen in sera of patients with pulmonary tuberculosis was $0.228{\pm}0.167$ in minimal tuberculosis; moderately advanced, $0.556{\pm}0.616$; far advanced, $1.116{\pm}0.651$ and $0.315{\pm}0.245$ in miliary tuberculosis. 2. The activity level (O.D. at 492nm) of IgG to BE antigen in sera of patients with pulmonary tuberculosis was $0.190{\pm}0.162$ in minimal tuberculosis; moderately advanced, $0.337{\pm}0.361$; far advanced, $0.713[\pm}0.460$ and $0.204{\pm}0.162$ in miliary tuberculosis. 3. The activity level (O.D. at 492nm) of IgG to AE antigen in sera of patients with pulmonary tuberculosis was $0.165{\pm}0.114$ in minimal tuberculosis; moderately advanced, $0.392{\pm}0.494$; far advenced, $0.751{\pm}0.512$ and $0.233{\pm}0.191$ in miliary tuberculosis. 4. The activity level (O.D. at 492nm) of IgG to FE antigen in sera of patients with pulmonary tuberculosis was $0.280{\pm}0.227$ in minimal tuberculosis; moderately advanced, $0.460{\pm}0.564$ ; far advanced, $0.845{\pm}0.573$ and $0.257{\pm}0.103$ in miliary tuberculosis. 5. The activity level (O.D. at 492nm) of IgG in sera of healthy control person was $0.126{\pm}0.084$ to TE antigen. $0.105{\pm}0.041$ to BE antigen, $0.103{\pm}0.052$ to AE antigen, and $0.095{\pm}0.061$ to FE antigen. 6. Degree of correlation(r) in activity level of IgG between TE antigen and BE antigen was 0.905 ; between TE antigen and AE antigen, 0.760; between TE antigen and FE antigen, 0.790, and between AE antigen and FE antigen, 0.945. 7. As O.D. above 0.200 was determined positive for the serodiagnosis of pulmonary tuberculosis, the sensitivity and specificity in ELISA using TE antigen were 80% and 87% respectively, whereas in the case of using BE antigen, 66% and 100%; in the case of using AE antigen, 62% and 100%, and in the case of using FE antigen, 72% and 93%, respecitively.

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Ultrasensitive Enzymeimmunoassay for Testosterone in Human Saliva (사람 타액내 Testosterone의 초감도 효소면역측정법)

  • 윤용달;전은현;이창주;도병록;이준영
    • Development and Reproduction
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    • v.4 no.1
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    • pp.115-123
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    • 2000
  • A few enzymeimmunoassay (EfA) for testosterone (T) have been reported but was not suitable for all biological samples. The present study was designed to develop a rapid, ultrasensitive EIA and to apply this technique for study the physiological changes of T in biological samples. Saliva samples were collected at 06:00~09:00 hour during one menstrual cycle from 18 normally menstruating women and on 09:00~10:00 hour from 20 normal men. The present study shows an established EIA for testosterone, using horseradish peroxidase (HRP), which was covalently bonded to testosterone-3-carboxymethyloxime (T-3-CMO). One batch of T-antisera was also covalently linked to microcrystalline cellulose particles by a mixed anhydride method in order to facilitate separation of bound and free steroids. The established EIA was validated in terms of sensitivity, accuracy, specificity, precisions etc., comparing with conventional radioimmunoassay. The sensitivity of the established EIA was less than 25 pg/tube. The correlation coefficients between the expected T-values and observed T-values measured by EIA or RIA were r=0.985 and r=0.941 respectively. The cross reactivity of antiserum in EIA was a little higher than that of RIA, especially by 5 ${\alpha}$-DHT. The intra- and inter-assay precisions of the present EIA were similar to those of RIA. The present study also demonstrates that the normal T-values in saliva of Korean male & female samples are 265.65${\pm}$15.80 pmol/l and 109.74${\pm}$ 12.01 pmol/l, respectively. The present EIA seems to be established and suitable for use in the endocrinological studies. The advantages of this EIA system also might make the present T-EIA an ideal procedure for use in a routine assay of ordinary laboratory with a conventional spectrophotometer.

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Studies on the Seroepidemiology of Helminthic Diseases in Korea (우리나라의 주요 기생충질환(寄生蟲疾患)에 대한 혈청역학적(血淸疫學的) 조사(調査))

  • Rim, Han-Jong;Lee, Joon-Sang;Joo, Kyoung-Hwan;Chung, Myung-Sook
    • Journal of agricultural medicine and community health
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    • v.16 no.1
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    • pp.48-60
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    • 1991
  • In a seroepidemiological study in several areas of Korea, the ELISA technique was performed to determine prevalence of some important helminthic diseases in our nation during March $15^{th}$ to June $30^{th}$. 1991. In this survey the serum antibody positive rates of anisakiasis, toxocariasis, clonorchiasis, paragonimiasis, cysticercosis, and sparganosis were measured. Among, 6,704 cases examined, 19.7% showed positive antibody titer at least one of the six items studied. Overall positive antibody rate was 8.1% in anisakiasis, 5.6% in toxocariasis, 3.6% in clonorchiasis, 1.7% in paragonimiasis, 4.5% in cysticercosis, and 2.6% in sparganosis respectively. In Pusan port southeastern part of Korea, antibody positive rate of anisakiasis was 2.9%, and clonorchiasis was 2.8% among 450 examine. In TaeJ$\check{o}$n city, central part of Korea. toxocariasis(6.7%) and anisakiasis(3.7%) showed high serologic positive rate. Of the 875 persons in Chunche$\check{o}$n gun(=province), northern central rural area of South Korea, anisakiasis was revealed as 3.4% seropositivity. In Tonghae port, eastern coast of South Korea. 9.9% of population examined showed positive antibody titer in anisakiasis. Of the 1,122 persons examined in Southern part of Cholla-Namdo(Southwestern coastal area of Korea), anisakiasis was 16.9%, cysticerocosis was 12.7% and the paragonimiasis was 3.3% respectively. In some localized area of Cholla-Pukdo, anisakiasis was 9.3% and cysticekosis was 4.3% among 702 cases examined. In some localized area of Kyungsang-Pukdo, anisakiasis was 10.6%. and toxocariasis was 16.1% among 900 cases examined. And finally, in Cheju-do, southern island of Korea, anisakiasis showed high positive rate(6.7%). Because cross reactions between related helminth group may disturb the analysis of these data, use of further developed techniques such as EITB(enzyme-linked immunoelectrotransfer blot) was considered as a essential tools for the study. We thought that probably most of the positive cases of cysticerosis were taeniasis cases. We can't rule out taeniasis even though EITB was employed as far as crude worm extract or cystic fluid of cysticercus was used as antigen. It was well Known that toxocariasis and anisakiasis also showed cross reactivity. However, the data presented here focus on seropositive rate of several helminthic diseases in Korea, not true prevalence rate of helminthiases, and to wait for more expensive purified antigen in sufficient amount for epidemiologic use is not necessary because increased immunologic sensitivity had little effect on epidemiologic sensitivity. We, here, suggest that ELISA should be applied as soon as possible to the evaluation of prevalence of tissue invading parasitic diseases, and a review of the antibody positive rate obtained in this study would be a basic data for controlling program of parasitic diseases in Korea.

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A Study on the Fouling of Ultrafiltration Membranes Used in the Treatment of an Acidic Solution in a Circular Cross-flow Filtration Bench (순환식 막 모듈 여과장치를 이용한 산성용액의 수처리 공정 시 발생하는 한외여과막 오염에 관한 연구)

  • Kim, Nam-Joon;Choi, Chang-Min;Choi, Yong-Hun;Lee, Jun-Ho;Kim, Hwan-Jin;Park, Byung-Jae;Joo, Young-Kil;Kang, Jin-Seok;Paik, Youn-Kee
    • Membrane Journal
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    • v.19 no.3
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    • pp.252-260
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    • 2009
  • The effects of the treatment of an acidic solution at pH 2 on polyacrylonitrile ultrafiltration (UF) membranes were investigated using a circular cross-flow filtration bench with a membrane module. A substantial reduction in the membrane permeability was observed after 80 hours' treatment of the acidic solution. In addition, the analyses of the sample solutions by ultraviolet/visible absorption spectroscopy and gas chromatography/mass spectrometry (GC/MS), which were taken from the feed tank as a function of the treatment time, showed that a new organic compound was produced in the course of the treatment. From a thorough search of the mass spectral library we presumed the new compound to be 1,6-dioxacyclododecane-7,12-dione (DCD), one of the well-known additives for polyurethane. Based on further experimental results, including the scanning electron microscope (SEM) images and the solid-state NMR spectra of the membranes used for the treatment of the acidic solution, we suggested that the decrease of the permeate flux resulted not from the deformation of the membranes, but from the fouling by DCD eluted from the polyurethane tubes in the filtration bench during the treatment. Those results imply that the reactivity to an acidic solution of the parts comprising the filtration bench is as important as that of the membranes themselves for effective treatments of acidic solutions, for efficient chemical cleaning by strong acids, and also in determining the pH limit of the solutions that can be treated by the membranes.

Pseudomonas tolaasii bacteriophage-specific polyclonal antibody formation and its cross reactivity to various phages (Pseudomonas tolaasii 박테리오파지에 특이적인 다클론항체 형성 및 이를 이용한 파지 교차 반응성)

  • Yun, Yeong-Bae;Park, Soo-Jin;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
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    • v.62 no.3
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    • pp.287-292
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    • 2019
  • Pseudomonas tolaasii causes brown blotch disease on the oyster mushroom (Pleurotus ostreatus). Various pathogenic strains of P. tolaasii were isolated and divided into three subtypes, $P1{\alpha}$, $P1{\beta}$, and $P1{\gamma}$. For phage therapy, bacteriophages against to these subtype strains were applied to mushroom cultivation and very successful to prevent from the disease. In this study, bacteriophages were isolated against the representative strains of subtype pathogens and their polyclonal antibodies were synthesized to investigate structural relationship among capsid proteins of phages. Phage preparations over $10^{10}pfu/mL$ were injected to rabbit thigh muscle and polyclonal antibodies were obtained after three times of boost injection. Titers of the antibodies obtained were over $2{\times}10^7Ab/mL$ for the phage ${\phi}6264$, $1{\times}10^6Ab/mL$ for the phage ${\phi}HK2$, and $1{\times}10^7Ab/mL$ for the phage ${\phi}HK19$ and phage ${\phi}HK23$. High specific activities were observed between antibodies and the corresponding bacteriophages. Some cross-reactivities between the antibodies and non-corresponding bacteriophages were also measured. Antibody $Ab{\phi}6264$ inactivated all phages of $P1{\alpha}$ subtype and only phage ${\phi}HK16$ among $P1{\beta}$ subtype phages. Antibody $Ab{\phi}HK23$ of $P1{\gamma}$ subtype neutralized all phages of $P1{\beta}$ subtype as well as the phage ${\phi}HK23$, showing the widest phage-inactivation range. When the structural-similarity studies of phages were investigated by using phage antibodies, closeness obtained by phylogenetic analysis of 16S rRNA genes of pathogenic strains were quite different from that of polyclonal antibody-specific structural similarity of phage capsid proteins. In conclusion, there is weak correlation between the host strain specificity of bacteriophage and its capsid structural similarity measured by phage antibodies.

Changes in Plasma and Urine Endothelin Levels During Acute Exacerbation of Asthma (급성 천식발작시 혈장 및 요중 Endothelin 농도 변동)

  • Chang, Jung-Hyun;Shin, Tae-Rim;Woo, Ga-Eun;Kim, Jong-Seon;Hong, Eun-Soon;Seo, Gi-Yeoul;Cha, Joo-Hyun;Kim, Mi-Seon;Kim, Yeung-Seon;Cho, Young-Joo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.844-852
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    • 1997
  • Background : Recent studies have documented increased release of endothelin(ET) during acute attack of asthma. The purpose of this study is to observe the link between plasma level and urinary excretion of each and changes during acute exacerbation. Method : Plasma and 24 hour urine were collected from sixteen asthmatics during acute exacerbation, twice ; first day of symptomatic exacerbation and two weeks after treatment. Controls were ten healthy normal subjects. All patients were treated with corticosteroid and beta-2 adrenergic agonist on admission. ET was determined by radioimmmunoassay and had 100% cross reactivity with ET-1, 67% with ET-2, 84% with ET-3, and 8% with Big-ET. Results : Plasma ETs were significantly elevated during acute attack of asthma compared with those in remission and controls.,However, there was no significant changes in urine ET concentrations or total ET amounts in 24 hour urine during exacerbation upto two weeks. Those levels of urine ET in asthmatics were still higher than controls. ET concentrations in plasma or urine were not correlated with pulmonary functional parameters and hypoxemia. Conclusion : The findings suggests that increased plasma ETs are related with exaggerated release during acute asthma. Urinary ET excretion is increased in asthma. However, urine ET changes during exacerbation should be observed in a larger and longer scale.

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Studies on the Immunodiagnosis of Rabbit Clonorchiasis 2. Immunoamnity purification of whole worm antigen and characterization of egg, metacercaria and adult antigens of Clonorchis sinensis (간흡충 감염 가토의 면역진단에 대한 연구 2. 성충 조항원의 정제 및 발육단계별 항원 분석)

  • Lee, Ok-Ran;Jeong, Pyeong-Rim;Nam, Hae-Seon
    • Parasites, Hosts and Diseases
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    • v.26 no.2
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    • pp.73-86
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    • 1988
  • The sensitivity and specificity of crude and affinity-purified antigens of Clcnorchis sinensis obtained from the infected rabbits were studied. Stage-specific antigenic proteins from the eggs, metacercariae and adult worms were characterized by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunosorbent astray (ELISA). The results were as follows: 1. The antibody.binding antigen (ABA) purified from whole worm crude antigen (IVWA) by CNBr-activated Sepharose 4B affinity chromatography made :l specific bands against rabbit antisera on Ouchterlony gel diffusion plate, while WWA made 7 bands. Major WWA protein bands by SDS-PAGE were found at 16, 300~18, 500 and 28, 000~29, 000 daltons, while major ABA protein bands were at 18, 000~21, 000 and 29, 000~31, 000 daltons. The reactivity of ABA with rabbit anti-sera in ELISA was remarkably less sensitive than that of WWA. 2. Molecular weights of egg antigen (EGA), metacercarial antigen (MEA) and adult worm antigen (WWA) of C. sinensis ranged from 15, 000-200, 000 daltons, 15, 000-100, 000 daltons and 11, 000~80, 000 daltons, respectively. Major WWA proteins consisted mainly of polypeptide bands of low molecular weight, less than 31, 000 daltons, while those of EGA and MEA consisted of higher molecular T.eights than 30, 000 daltons. 3. The ELISA reactivities of WWA to rabbit anti.sera were remarkably greater than those of MEA. EGA showed negative reaction throughout the experiments. WWA showed higher optical density (O.D.) than 1.0, when reacted with rabbit anti-sera obtained at 4~6 weeks after the infection. In the rabbit anti-sera later than 12 weeks after the infection, the O.D. reacting witll WWA showed a plateau without variation. MEA shoT.ed relatively low O.D. values (<0.6), when reacted with anti-sera from lightly in(ected groups throughout the experiments, althougll there were some wealth positive cases (O.D.>0.6) ill heavily infected groups. MEA reacted with rabbit anti-sera showed negative results on Ouchterlony gel diffusion plates. Summarizing the above results, it is suggested that the whole worm antigen prepared from the adult worms of C. sinensis is most highly antigenic. However, this antigen might reveal cross reactions with other trematodes such as Paragonimus westermani, therefore, purification of antigenic proteins from the crude antigen is essential 18 increase the sensitivity and specificity for the immuncdiagnosis of clonorchiasis.

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