• Title/Summary/Keyword: Chemical antigen

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Serodiagnosis of canine toxoplasmosis by latex agglutination and indirect fluorescent antibody test (Latex 응집반응과 간접형광항체법을 이용한 개 톡소플라즈마병의 혈청학적 진단)

  • Lee, Byung-hoon;Lee, Eung-goo;Suh, Myung-deuk
    • Korean Journal of Veterinary Research
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    • v.32 no.4
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    • pp.641-647
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    • 1992
  • This study was conducted to determine the serum antibodies against toxoplasma in the artificially infected dogs, pet and street dogs by latex agglutination (LA) and indirect fluorescent antibody (IFA) test. LA test was carried out with commercial Toxo-MT kit (Eiken chemical Co.) and IFA test was carried out with rabbit-anti-dog IgG labelled with FITC (Cappel Co.) and toxo-antigen slides prepared in laboratory. The results obtained were as follows ; 1. Antibodies to Toxoplasma gondii in the artificially infected dogs were detected firstly at the Day 8 in IFA and Day 9 in LA test after inoculation. Positive antibody reactions by these tests were declined gradually afterward but maintained up to 12 weeks. 2. In LA test serum antibody titers in 310 test sera were shown as 10 cases(32%) in 1 : 32.5(1.0%) in 1 : 64, 4(1.3%) in 1 : 128 and 2(0.7%) in 1 : 256. In IFA test serum antibody titers 310 test sera were shown as 17 cases(5.5%) in 1 : 64, 8(2.6%) in 1 : 128 and 5(1.6%) in 1 : 256. 3. In the total of 310 sera from pet and street dogs toxoplasma antibody positive rates were 21 cases (6.8%) by LA and 30 cases (9.7%) by IFA test and the positive detection rates between these two groups by LA and IFA test were not significant(p<0.05). 4. In the total of 115 sera from pet dogs toxoplasma antibody positive rates were 12 cases(10.4%) by LA and 15(13.0%) by IFA test. And in the 195 street dogs the positive rates were 9 cases(4.6%) by LA and 15(7.7%) by IFA test. Also, the positive detection rates between these two groups by LA and IFA test were not significant(p<0.05). 5. Agreement of reactivity between LA and IFA test for 310 sera was 91.3% in total of 283 cases consisting of 12 cases(3.9%) of both LA and IFA positive and 271 cases(87.4%) of LA and IFA negative. 6. LA test was almostly equivalent to the IFA test in producibility and proved to be a simple tool for the screening of toxoplasma antibody in laboratory.

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Studies on the Rapid Plasma Reagin(RPR) Card Test for the Diagnosis of Syphilis (매독진단(梅毒診斷)을 위(爲)한 신속(迅速)한 혈장항체검사법(血漿抗體檢査法)에 관(關)한 연구(硏究))

  • Kim, Joo-Deuk;Lew, Joon;Kim, Hyun-Joo
    • The Journal of the Korean Society for Microbiology
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    • v.3 no.1
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    • pp.15-23
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    • 1968
  • For the effective control of Syphilis, many investigators have developed a more rapid, simple and economical screening serological test which is adequately sensitive and specific. To fulfill the requirements of a more rapid serologic test for syphlis, a substitute for the conventional serum specimen was needed since considerable time and labor are involved in the processing of blood to serum. Burdon suggested the use of plasma in the serologic tests for syphilis as a substitute for serum. He noticed that plasma was more sensitive than serum in the Kline and Kahn tests, and attributed this to the presence of more antibody-like substance, "reagin" in plasma than in serum. However, to make plasma sufficiently sensitive, it was necessary to inactivate plasma by heating at a temperature of $56^{\circ}C$ for about 30 minutes. Heating of plasma resulted in the precipitation of fibrinogen which made centrifugation necessary to obtain dear plasma. Since the chief disadvantage to the use of unheated plasma(or serum) was a reduction in sensitivity of results-which probably was due to a labile factor such as complement-Portnoy et al began to consider rapid chemical methods of inactivation of plasma(or serum). They experienced that choline chloirde was shown to be anticomplementary which suggested its use as an inactivating agent for unheated plasma(or serum). In 1959 Portnoy et al reported the Rapid Plasma Reagin(RPR) Test for syphilis which is a more rapid, economical and simple. But still this test has many disadvantages as a rapid performing, field and office procedure, because it requires the usual laboratory equipments such as centrifuge, rotating machine, microscope etc. To substitute these disadvantages of the RPR test, in 1962, Portnoy et al developed the Rapid Plasma Reagin(RPR) card test for syphilis, which has the following advantages: a) Simplicity and rapidity of performance, b) Requires no laboratory equipments, c) Stable antigen suspension, d) Adequate sensitivity and specificity. This RPR card test can be used as a rapidly performing and screening test in field investigation, outpatient clinics, small laboratories and hospitals doing limited syphilis serology, and predonor in blood bank. Private clinic which has limited laboratory equipment and technic for syphilis serology can also use this RPR card test as a tool in the rapid diagnosis of syphilis. It was thought that this RPR card test is a useful tool in Korea for private physician and mass survey for syphilis diagnosis. But Portnoy patented the reagents needed for the performing the RPR card test. Therefore authors developed newly the reagents and according to Portnoy's method evaluated the newly developed. RPR card test compared with the VDRL, Kolmer CF, and RPCF tests. The RPR card and VDRL tests were performed plasma and serum from the total 1,132 cases. Among these 1,131 cases, 521 were syphilis suspected laboratory specimens, and 611 were syphilis unsuspected healthy young men. After screening with these two tests, the RPR card and VDRL tests, reactive specimens to the above one or both tests were retested by the Kolmer CF and RPCF tests.

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A Research and Application of Polyhydroxyalkanoates in Biosensor Chip (생분해성 고분자, 폴리하이드록시알카노에이트를 이용한 바이오센서 칩 연구와 그 응용)

  • Park, T.J.;Lee, S.Y.
    • KSBB Journal
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    • v.22 no.6
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    • pp.371-377
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    • 2007
  • Polyhydroxyalkanoates (PHAs) are a family of microbial polyesters that can be produced by fermentation from renewable resources. PHAs can be used as completely biodegradable plastics or elastomers. In this paper, novel applications of PHAs in biosensor are described. A general platform technology was developed by using the substrate binding domain (SBD) of PHA depolymerase as a fusion partner to immobilize proteins of interest on PHA surface. It could be shown that the proteins fused to the SBD of PHA depolymerase could be specifically immobilized onto PHA film, PHA microbead, and microcontact printed PHA surface. We review the results obtained for monitoring the specific interaction between the SBO and PHA by using enhanced green fluorescent protein, red fluorescent protein, single chain antibody against hepatitis B virus preS2 surface protein and severe acute respiratory syndrome coronavirus surface antigen as model proteins. Thus, this system can be efficiently used for studying protein-protein and possibly protein-biomolecule interactions for various biotechnological applications.

Fabrication of Label-Free Biochips Based on Localized Surface Plasmon Resonance (LSPR) and Its Application to Biosensors (국소 표면 플라즈몬 공명 (LSPR) 기반 비표지 바이오칩 제작 및 바이오센서로의 응용)

  • Kim, Do-Kyun;Park, Tae-Jung;Lee, Sang-Yup
    • KSBB Journal
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    • v.24 no.1
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    • pp.1-8
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    • 2009
  • In the past decade, we have observed rapid advances in the development of biochips in many fields including medical and environmental monitoring. Biochip experiments involve immobilizing a ligand on a solid substrate surface, and monitoring its interaction with an analyte in a sample solution. Metal nanoparticles can display extinction bands on their surfaces. These charge density oscillations are simply known as the localized surface plasmon resonance (LSPR). The high sensitivity of LSPR has been utilized to design biochips for the label-free detection of biomolecular interactions with various ligands. LSPR-based optical biochips and biosensors are easy to fabricate, and the apparatus cost for the evaluation of optical characteristics is lower than that for the conventional surface plasmon resonance apparatus. Furthermore, the operation procedure has become more convenient as it does not require labeling procedure. In this paper, we review the recent advances in LSPR research and also describe the LSPR-based optical biosensor constructed with a core-shell dielectric nanoparticle biochip for its application to label-free biomolecular detections such as antigen-antibody interaction.

Preparation of $^{125}I-Iodotyraminehemisuccinyltaxol\;(^{125}ITHT)$ for Competitive Taxol Radioimmunoassay (Taxol 방사면역측정을 위한 $^{125}I-Iodotyraminehemisuccinyltaxol\;(^{125}ITHT)$의 제조)

  • Lee, Tae-Sup;Awh, Ok-Doo;Choi, Tae-Hyun;Kim, Hyun-Suk;Hong, Jun-Pyo;Lee, Eun-Sook;Choi, Chang-Woon;Lim, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.36 no.2
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    • pp.121-132
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    • 2002
  • Purpose : Taxol(Paclitaxel), an antineoplastic agent, has been used in the treatment of ovarian and breast cancers. The determination of optimal Taxol concentrations in human serum was required for enhancing therapeutic effect and maintaining the appropriate Taxol level in blood. This study was aimed to synthesizeradiolabeled Taxol derivatives as radiotracer in competitive radioimmunoassay for monitoring Taxol concentrations in blood and to determine the usefulness of its derivatives. Materials and Methods : Hemisucdcinyltaxol(HT) was synthesized by esterification of Taxol with succinic anhydride. Tyraminehemisuccinyltaxol(THT) was synthesized by coupling of HT with tyramine using isobutylchlormate as coupling agent and purified by HPLC. By using chloramine-T($5.25mg/ml,\;10{\mu}{\ell}$) as oxidant agent, THT($4mg/ml,\;30{\mu}{\ell}$) was labeled wity $^{125}I\;(37MBq,\;1mCi)$. To estimate the stability of purified THT, $^{125}I-iodotyraminehemisuccinyltaxol(^125}ITHT)$ was dissolved in 80% acetonitrile aqueous solution, and the solution was incubated at $4^{\circ}C\;and\;37^{\circ}C$ for 7 days. At various time intervals, the stability of THT and $^{125}ITHT$ was monitored. The titer of Taxol monoclonal antibody, 3G5A7, was determined by competitive radioimmunoassay using $^{125}ITHT$ as a labeled antigen. A standard dose-response curve was demonstated by Taxol competitive radioimmunoassay. Resulls : HT and THT were synthesized with 79.9% and 19.5% yield, respectively. The labeling yield of $^{125}ITHT$ was 93%. After 7 days, the chemical purity of THT was 96.5% at $4^{\circ}C$, and 97.5% at $37^{\circ}C$. After 3 days, $^{125}ITHT$ was stable with 94.7% at $4^{\circ}C$ and 93.4% at $37^{\circ}C$. After 7 days, fadiochemical purity was diminished to 88.1% at $37^{\circ}C$. The titer of Taxol monoclonal antibody, 3G5A7, was determined to 1:256. A standard dose-response curve demonstated good collinearity ($R^2=0.971$) as Taxol concentration-dependent manner. Conclusion : Competitive radioimmunoassay using $^{125}I-iodotyraminehemisuccinytaxol$ as radiotracer could be used to monitor for concentration of Taxol in the human serum.