• 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.

• Tuberculosis and Respiratory Diseases
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• v.46 no.4
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• pp.473-480
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• 1999

Background: Early diagnosis of tuberculosis is critical, especially in Korea, an area where tuberculosis is endemic. Because antibody responses to some membrane proteins of Mycobacterium tuberculosis are not comparable, and the policy of BCG vaccination and the prevalence of tuberculosis are different from country to country, the usefulness of the serological diagnostic tests is questionable in Korea, even though they have been confirmed to be useful in other countries. In the specific context of Korea, we tried to evaluate the validity of the ICT Tuberculosis Test (ICT), a membrane-based antibody kit that purports to detect the 5 M. tuberculosis complex-specific antigens including 38-kDa protein. Method: 68 patients with tuberculosis were tested : 37 had no history of previous tuberculosis, and 31 were reactivated cases. The control group comprised 77 subjects : 25 healthy adults, 35 hospital workers with frequent contact with tuberculosis patients, and 17 in-patients with non-tuberculous respiratory diseases. Results: The diagnostic sensitivities of the ICT were 87% and 73% in patients with versus without previous history of tuberculosis, respectively. The sensitivities of smear-positive and smear-negative patient groups were 81% and 73%, respectively. Both of the two patients with extrapulmonary tuberculosis tested positive through the ICT. The specificities of the ICT were 88%, 94%, and 94% in healthy adults, hospital workers, and non-tuberculous patients, respectively, with an overall specificity of 92%. Conclusion: It is suggested that when combined with traditional techniques, the ICT is an useful tool for the diagnosis of pulmonary tuberculosis. The procedure is simple, easy to perform, rapid, and needs no equipment. It shows 73% sensitivity and 92% specificity for the diagnosis of tuberculosis.

• Parasites, Hosts and Diseases
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• v.25 no.2
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• pp.159-167
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• 1987

To analyse the antigen specificity of patients sera from 24 confirmed neurocysticercosis and a monoclonal antibody, SDS-PAGE using 10~15% linear gradient gel and EITB were done. Cystic fluid, saline extracts of scolex and of whole worm of C. cellulosae, saline extracts of sparganum, hydatid cyst fluid, saline extracts of Fasciola, Clonorchis and Paragonimus were used as antigen. Of protein bands in cystic fluid of C. cellulosae, patient sera reacted frequently to bands of 152, 94, 64, 48, 24, 15, 10 and 7kDa proteins. To saline extracts of scolex and whole worm of C. cellulosae, patients sera reacted frequently to 94, 64, 52, 39, 34, 15 and 10kDa bands. Two bands in sparganum extract (130 and 64kDa) and two bands in hydatid cyst fluid (52 and 27kDa) were cross-reacting bands with sera from cysticercosis patients. Saline extracts of Fasciola, ClonorchiJ and Paragonimus did 'not exhibit cross-reacting bands. Monoclonal antibody to cystic fluid of C. cellulosae was found to react with low molecular weight proteins of 15, 10 and 7kDa.

• Parasites, Hosts and Diseases
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• v.24 no.1
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• pp.25-41
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• 1986

The applicability of micro-ELISA was evaluated in human neuro-cysticercosis using paired samples of serum and CSF. A total of 355 cases who were mostly neurologic patients was subjected. Cystic fluid of C. cellulosae was used as antigen in protein concentration of $2.5{\;}{\mu}g/ml$. Serum was diluted to 1 : 100 and CSF was undiluted in the assay for the specific IgG antibody level. The differential criterion of the positive reaction was the abs. of o. 18 in both samples. The results were summarized as follows: 1. The overall sensitivity of the micro-ELISA in 71 confirmed neurocysticercosis was 90.1% ; the sensitivity by serum was 77.5% and that by CSF was 83.1%. CSF was a more sensitive and valuable material. Most of the false negative cases of neuro-cysticercosis showed far lower level of abs. rather than marginal. 2. The overall specificity of the micro-ELISA in 52 confirmed other neurologic diseases was 88.5%; the specificities by serum and by CSF were 94.2% respectively. Cases of other neurologic diseases did not show false positive reactions in both samples. 3. When serum was assayed, taeniasis(2/18), sparganosis(2/20), paragonimiasis(1/56), clonorchiasis(1/15) and fascioliasis(1/1) cases showed cross reactions. When CSF was assayed, 2 ot 10 neuro-sparganosis showed cross reactions while none of 9 neuro-paragonimiasis showed it. Out of 71 confirmed neuro-cysticercosis cases, 6 and 11 showed cross reactions by serum and CSF to crude extract antigen of sparganum; but no case did show it to crude extract antigen of Paragonimus westermani. 4. Ventricular CSF showed low or negative levels of IgG antibody than lumbar CSF unless the lesion was at the lateral ventricle itself. 5. Out of 4 racemose cysticercosis cases, 3 showed positive reaction in serum while all of 3 examined CSF were positive. The above results indicated that the serological test for detecting the specific IgG antibody by micro-ELISA using paired samples of serum and CSF was very helpful for clinical differentiation of neuro-cysticercosis from neurologic diseases of other causes.

• Parasites, Hosts and Diseases
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• v.24 no.2
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• pp.145-158
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• 1986

This study was undertaken to purify cystic fluid (CF) antigen of Taenia solium metacestodes by affinity chromatogaphy using specific monoclonal antibody(McAb) and to characterize the antigenicity of the purified antigen. The hybridoma cell lines, prepared by fusion between mouse plasmacytoma and spleen cells from BALB/c mice immunized with CF, secreted antibodies reacting to various helminthic antigens. Majority of cell lines reacted to CF only but some also reacted to parenchymal antigen of T. solium metacestodes, adult T. saginata, sparganum, hydatid cystic fluid, Paragonimus westermani and Clonorchis sinensis, either in combination with CF, other antigens or independently. Cloned cells derived from monoclonal lines also produced antibodies reacting either to CF only or to other helminthes in combination or independently. These results indicated that CF of T. solium metacestodes contained proteins which possessed antigenic determinants not only specific to CF but also cross reactive with the afore-mentioned helminthes. CF of T. solium metacestodes was purified by affinity chromatography using the McAb which reacted to CF and parenchymal antigens. The affinity-purified antigen (A-Ag) and unbound pool CF (U-Ag) were separated. A-Ag showed 2 protein bands by disc-PAGE whereas CF exhibited 6 bands and U-Ag consisted of all bands CF had. The diagnostic significance of A-Ag was evaluated by ELISA in human neurocysticercosis and other helminthic and neurologic diseases. By A-Ag, the levels of the specific IgG antibody, as shown by absorbance in sera and CSF, were lower than those of CF and U-Ag. Accordingly, the sensitivity was about 70% of CF and U-Ag. However, the nonspecific positive reactions to CF and U-Ag, observed in sparganosis, T. saginata infection and paragonimiasis did not occur when A-Ag was used. These results indicated that the affinity-purified A-Ag had the higher specificity but the lower sensitivity as a diagnostic antigen in cysticercosis, probably because it only detected a single or limited numbers of monospecific antibodies among the diverse polyclonal antibodies produced in the patients with neurocysticercosis.

• The korean journal of orthodontics
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• v.24 no.2
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• pp.303-317
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• 1994

This study was designed to measure the changes in the titer of tooth root antibodies accompanying root resorption associated with orthodontic tooth movement in dogs to explore a role of the specific immune response in root resorption during orthodontic tooth movement. Five adult mongrel dogs, 2 years of age, were used in the study. Six lower incisors were extracted as sources of homologous antigen in the dogs. Tooth root antigen preparations were made from a 6M Guanidine-HCl-10% EDTA(pH5.0) extract of these root dentins. Root resorption was elicited by intrusion of six maxillary incisors with 200-250gm intrusive force. In 9th week, resorbing six maxillary anterior teeth were extracted. Serum samples were taken from each dog prior to intrusion and weekly for 11 consecutive weeks. Serum autoantibody titers were determined with an enzyme-linked immunosorbent assay. As controls for antibody specificity, sera which were previously incubated with tooth root antigen as well as sera to an unrelated bacterial antigen (Porphyromonas gingivalis 33277) for 3 hours at 25 were measured in all runs. Root resorption was monitored monthly using occlusal radiographs. And then root resorption patterns were observed with a zoom stereo microscope (Model SZH-121, Olympus optical Co. Ltd.). Incisors did not show clear radiographic evidence of significant and progressive root resorption, but periodontal ligament space had widened. But root resorption was observed on the apical regions of the maxillary incisors with a zoom stereo microscope. Teeth showed the shallow depression generally accompanying deep resorption. These demonstrate a slight tendency for an immediate decrease followed by rebound to levels above the pre-treatment baseline. A peak titer of autoantibody to dentin antigen occurred on day 28, then steadily decreased during the 9th week period as the roots resorbed and then rapidly spiked in animals when the resorbing teeth were extracted. When sera is incubated with tooth root antigen, serum activity in the ELISA was almost absent. This is because serum activity in the ELISA could be removed by absorption of the serum with dog dentin antigen. Serum ELISA activity to the unrelated bacterial antigen remained essentially unchanged in all animals throughout the experimental period. When the time course of changes in autoantibody to homologous tooth root antigen prepatration and unrelated bacterial antigen was compared, no significant differences were found(${\alpha}=0.05$). In general, the overall pattern of changes in autoantibody was similar to the two antigens. These findings suggest the possibility that these immunologic changes precede a significant development of root resorption lesions rather than merely reflecting their presence. Therefore, this suggests that the changes of antibody levels may have some predictive value for root resorption.

• Tuberculosis and Respiratory Diseases
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• v.44 no.4
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• pp.785-795
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• 1997

Background : Tumor markers have been used in diagnosis, predicting the extent of disease, monitoring recurrence after therapy and prediction of prognosis. But the utility of markers in lung cancer has been limited by low sensitivity and specificity. TPA-M is recently developed marker using combined monoclonal antibody of Cytokeratin 8, 18, and 19. This study was conducted to evaluate the efficacy of new tumor marker, TPA-M by comparing the estabilished markers SCC, CEA, Cyfra21-1 in lung cancer. Method : An immunoradiometric assay of serum CEA, sec, Cyfra21-1, and TPA-M was performed in 49 pathologically confirmed lung cancer patients who visited Keimyung University Hospital from April 1996 to August 1996, and 29 benign lung diseases. Commercially available kits, Ab bead CEA (Eiken) to CEA, SCC RIA BEAD (DAINABOT) to SCC, CA2H (TFB) to Cyfra2H. and TPA-M (DAIICHI) to TPA-M were used for this study. Results : The mean serum values of lung cancer group and control group were $10.05{\pm}38.39{\mu}/L$, $1.59{\pm}0.94{\mu}/L$ in CEA, $3.04{\pm}5.79{\mu}/L$, $1.58{\pm}2.85{\mu}/L$ in SCC, $8.27{\pm}11.96{\mu}/L$, $1.77{\pm}2.72{\mu}/L$ in Cyfra21-1, and $132.02{\pm}209.35\;U/L$, $45.86{\pm}75.86\;U/L$ in TPA-M respectively. Serum values of Cyfra21-1 and TPA-M in lung cancer group were higher than control group (p<0.05). Using cutoff value recommended by the manufactures, that is $2.5{\mu}/L$ in CEA, $3.0{\mu}/L$ in Cyfra21-1, 70.0 U/L in TPA-M, and $2.0{\mu}/L$ in SCC, sensitivity and specificity of lung cancer were 33.3%, 78.6% in CEA, 50.0%, 89.7% in Cyfra21-1, 52.3%, 89.7% in TPA-M, 23.8%, 89.3% in SCC. Sensitivity and specificity of nonsmall cell lung cancer were 36.1%, 78.1% in CEA, 50.1%, 89.7% in Cyfra21-1, 53.1%, 89.7% in TPA-M, 33.8%, 89.3% in SCC. Sensitivity and specificity of small cell lung cancer were 25.0%, 78.5% in CEA, 50.0%, 89.6% in Cyfra21-1, 50.0%, 89.6% in TPA-M, 0%, 89.2% in SCC. Cutoff value according to ROC(Receiver operating characteristics) curve was $1.25{\mu}/L$ in CEA, $1.5{\mu}/L$ in Cyfra2-1, 35 U/L in TPA-M, $0.6{\mu}/L$ in SCC. With this cutoff value, sensitivity, specificity, accuracy and kappa index of Cyfra21-1 and TPA-M were better than CEA and SCC. SCC only was related with statistic significance to TNM stages, dividing to operable stages(TNM stage I to IIIA) and inoperable stages (IIIB and IV) (p<0.05). But no tumor markers showed any correlation with significance with tumor size(p>0.05). Conclusion : Serum TPA-M and Cyfra21-1 shows higher sensitivity and specificity than CEA and SCC in overall lung cancer and nonsmall cell lung cancer those were confirmed pathologically. SCC has higher specificity in nonsmall cell lung cancer. And the level of serum sec are signiticantly related with TNM staging.

• Pediatric Infection and Vaccine
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• v.6 no.1
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• pp.78-85
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• 1999

Purpose : Mycoplasma pneumoniae pneumonia has been to be developed frequently in school age children and adolescence and hard to see under 3 year-old children. But it seems to be increased in number of patients with Mycoplasma pneumoniae pneumonia under 3-year old in clinical practice in these days. We have aimed to examine the characteristics of clinical findings of Mycoplasma pneumonia under 3 year-old children. Methods : We had performed retrospective review of medical records of 30 patients with Mycoplasmal pneumonia under 3-year old children who admitted to Department of Pediatrics, Kyunghee University Hospital from Jan. 1994 to Dec. 1997. The diagnostic criteriae was Cold agglutinin titer>1:64 or Mycoplasma antibody titer>1:80. Results : Mycoplasmal pneumonia was 30 out of 235 cases(12.7%) of total pneumonia under 3 year old children. Male female ratio was 1.3 : 1 and age distributions were 0~1y : 0, 1~2y : 8, 2~3y : 22 cases. Clinical symptoms and signs were cough(100.0%), sputum(83.3%), fever(80.0%) rhinorrhea(33.3%), vomiting(33.3%), moist rale(86.7%), decreased breathing sound(26.7%), wheezing(20.0%), and pharyngeal injection(30.0%). Thirteen out of 30 cases(43.3%) had unilateral infiltration, 10 cases(33.4%) had bilateral infiltration, 1 case(3.3%) had pleural effusion, and 6 cases(20.0%) had negative findings on chest radiography and there was no cases of atelectasis. On laboratory findings, 6 out of 30 cases(20.0%) had leukocytosis, 1 case(3.3%) had neutrophilia, 10 cases(30.0%) had eosinophilia, 17 cases(56.7%) had increased ESR, and 18 cases(60.6%) had positive CRP. Positive cold agglutinin titers(>1 : 64) were 19 cases(63.3%), and positive mycoplasma antibody(M-ab) titers(>1 : 80) were 27 cases(93.3%). Mycoplasma antibody test was more valuable than cold agglutinin test for the diagnosis of Mycoplasmal pneumonia and there was no correlation between cold agglutinin titer and mycoplasma antibody titer. Mycoplasma-polymerase chain reaction(M-PCR) was done with 13 cases, 12 out of 13 cases(92.3%) were positive. M-PCR test was valuable to the diagnosis of Mycoplasmal pneumonia but it will be needed to further study for their clinical application. Among 30 cases, 5 cases(16.7%) had complications, 3 cases(10.0%) had skin rash, 1 case(3.3%) had pleural effusion, 1 case(3.3%) had arthralgia, but all complications were mild and recovered without residual sequelae. Conclusion : The occurrence of Mycoplasmal pneumonia under 3 year-old children was not rare from this study. Clinical characteristics of Mycoplasmal pneumonia under 3-year old were normal radiologic findings in many cases, low complication rate, mild clinical course, and tend to rapid recovery compared with general manifestations of Mycoplasmal infectionsin children and adolescence. There were likely to be missed patients with Mycoplasmal pneumonia which did not diagnose by conventional serologic tests that had low sensitivity and specificity. We have to pay attention to the Mycoplasmal infection of the young children with pneumonia during epidemic periods of Mycoplasmal infection.

• Parasites, Hosts and Diseases
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• v.50 no.1
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• pp.15-21
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• 2012

In Iran, $Plasmodium$ $vivax$ is responsible for more than 80% of the infected cases of malaria per year. Control interventions for vivax malaria in humans rely mainly on developed diagnostic methods. Recombinant $P.$ $vivax$ apical membrane antigen-1 (rPvAMA-1) has been reported to achieve designing rapid, sensitive, and specific molecular diagnosis. This study aimed to perform isolation and expression of a rPvAMA-1, derived from Iranian patients residing in an endemic area. Then, the diagnostic efficiency of the characterized Iranian PvAMA-1 was assessed using an indirect ELISA method. For this purpose, a partial region of AMA-1 gene was amplified, cloned, and expressed in pET32a plasmid. The recombinant $His-tagged$ protein was purified and used to coat the ELISA plate. Antibody detection was assessed by indirect ELISA using rPvAMA-1. The validity of the ELISA method for detection of anti-$P.$ $vivax$ antibodies in the field was compared to light microscopy on 84 confirmed $P.$ $vivax$ patients and compared to 84 non-$P.$ $vivax$ infected individuals. The ELISA cut-off value was calculated as the mean+2SD of OD values of the people living in malaria endemic areas from a south part of Iran. We found a cut-off point of OD=0.311 that showed the best correlation between the sera confirmed with $P.$ $vivax$ infection and healthy control sera. A sensitivity of 81.0% and specificity of 84.5% were found at this cut off titer. A good degree of statistical agreement was found between ELISA using rPvAMA-1 and light microscopy (0.827) by Kappa analysis.

• Korean Journal of Biological Psychiatry
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• v.2 no.1
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• pp.100-106
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• 1995

Antibodies to hepatitis C drew attention because of high morbidity to chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. HCV was known to be transmitted by transfusion, sexual behavior and parenteral drug use. However, some kind of autoimmune mechanism was suggested to be involved in the genesis of HCV-induced liver diseases. We hypothesized the prevalence of having anti-HCV might be higher in psychiatric patients rather than general population because of the characteristic route of transmission. Using Abbott HCV BA kit, anti-HCV was detected in the sera of 113 psychiatric inpatients from early December in 1992 to late May in 1994. The Positivity of anti-HCY was significantly(P<0.05) higher among psychiatric inpatients(10.6%) than in healthy controls(3.0%). There were no disease specificity among psychiatric inpatients who had anti-HCV, though alcoholics tended to have more anti-HCV. We couldn't find any significant correlation of anti-HCV with age, seasons of birth, lymphocytes (%) and liver function.