• Title, Summary, Keyword: chemiluminescence immunoassay

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Development of Chemiluminescence Immunoassay for Progesterone in Serum (혈청내의 Progesterone 측정을 위한 Chemiluminescence Immunoassay의 개발에 관한 연구)

  • Lee, K.S.;Suh, B.H.;Lee, J.H.;Kim, J.B.
    • Clinical and Experimental Reproductive Medicine
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    • v.17 no.1
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    • pp.87-91
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    • 1990
  • Development of a solid-phase chemiluminescence immunoassay for the detection of progesterone in serum extract was described. The chemiluminescence immunoassay was establised utilizing anti-progesterone monoclonal antibody that coated on polystyrene tubes and progesterone-ABEI conjugate as tracer. The light yield generated from antibody bound conjugate was counted on clinilumat luminometer by oxidation with microperoxidase and peroxide. The chemiluminescence immunoassay was high specific and accurate and detects as little as 3.9ng/ml of progesterone. The intra-assay CV ranged from 6% to 11.5% and inter-assay CV ranged from 13.6% to 18.7%. This assay system was good correlated with conventional kit radioimmunoassay system (r=0.98).

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Molecular Cloning of H-Y Antigen Gene I. Purification of H-Y Antigen by Immunoaffinity Chromatography and Chemiluminescence Immunoassay for the Assay of H-Y Antigen (H-Y 항원 유전자의 cloning에 관한 연구 I. 친화성 크로마토그래피에 의한 H-Y 항원의 분리 정제 및 H-Y 항원 정량을 위한 화학발광 면역 분석법)

  • 김종배;김재홍;백정미;김창규;정길생
    • Korean Journal of Animal Reproduction
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    • v.15 no.2
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    • pp.149-155
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    • 1991
  • 본 실험은 H-Y 항원 유전자 크로닝을 위한 기초연구로서 H-Y 항원의 특성을 규명하기 위하여 친화성 크로마토그래피에 의하여 H-Y 항원을 분리·정제하였다. 정소 추출액을 항체가 결합된 column에 결합시킨 뒤 10% acetic acid로 용출시켰다. 용출된 분획을 모아 농축한 후 HPLC와 SDS-PAGE를 실시하여 H-Y 항원의 분자량은 약 6,7000달톤 임을 알 수 있었으며 isoelectric focusing에 의하여 등전점(pI)은 5.0인 것으로 측정되었다. H-Y 항원에 대한 단일클론항체와 표지항원으로는 H-Y 항원-ABEI(aminobutylethyl isoluminol)를 사용하여 H-Y 항원 정량을 위한 화학발광면역분석법을 개발하였다. 항원항체 반응후 빛의 측정은 NaOH 존재하에서 microperoxidase/H2O2를 이용한 산화반응으로 실시하여 10초간 측정한 빛의 양을 적분하였다. H-Y 항원의 농도와 빛의 양과는 역비례하였으며 감도는 11.8ng/tube 정도이었다.

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Development of Chemiluminescence Immunoassay(CIA) & ELISA for the Detection of Anti-sperm Antibodies in Male Serum (항정자 항체 검출을 위한 CIA 및 ELISA 개발을 위한 기초 연구)

  • Kim, S.C.;Lee, K.S.;Kim, Y.K.;Kim, C.K.;Choi, K.H.;Kwon, O.J.;Kim, J.B.
    • Clinical and Experimental Reproductive Medicine
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    • v.17 no.1
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    • pp.71-80
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    • 1990
  • New immunoassay systems for the detection of anti-sperm antibodies were developed. For this, sperm surface protein was purified by the immunoaffinity column prepared by the coupling of rabbit anti-human IgG antibodies to Sepharose-4B. Fraction eluted by tris-HCI buffer containing SDS showed a single band having molecular weight of about 60KD on electrophoresis. Enzyme HRP labelled goat anti-human IgG and chemiluminescence aminobutylethyl-isoluminol(ABEI) labelled rabbit anti-human IgG were used for ELISA and CIA, respectively. These two labelled conjugate bound well with human IgG. When serum dilution curves were made to titrate positive serums, two kinds of curves with steep and sluggish slopes were obtained Serum samples were categorized into 3 groups: positive, weak positive and negative based on slope of curve and O.D. values at 1:160 dilution of serum. When ELISA and CIA were compared to conventional method Kibrick test by the determinations of 62 male serums with different diagnosis, the results of ELISA and CIA agreed well, but both disagreed with that of Kibrick test. This study showed that purified sperm surface antigen can be used to develope solid-phase immunoassay systems such as ELISA and CIA which may eliminate the problems encounted the immobilization of living sperm in other tests.

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Chemiluminescence Immunoassay for Measurement of Estrone-3-Sulfate Using Monoclonal Antibody to Estrone-3-Glucuronide (Estrone-3-Glucuronide에 대한 단일클론항체를 이용한 Estrone-3-Sulfate 측정을 위한 화학발광면역분석법)

  • 김윤규;민형식;김춘원;김창규;김선호;김종배
    • Korean Journal of Animal Reproduction
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    • v.20 no.2
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    • pp.215-221
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    • 1996
  • This study was carried out to develop an immunoassay for the diagnosis of the pregnancy and ovarian function of domestic animals. Using 2E92C10 monoclonal antibody(McAb) generated against estrone-3-glucuronide(E1-3-G) and appeared a high cross-reactivity with estrone-3-sulfate(E1-3-S), chemiluminesence immunoassay (CIA) to detect E1-3-S was developed. 2E92C10 McAb cross-reacted with E1-3-S (30%) was purified from ascites fluid using protein G sepharose gel column. The purity of purified antibody fraction was monitored by SDS-PAGE and was better compared to that of crude ascite fluid. The soild and liquid phase CIA for E1-3-S were established utilizing 2E92C10 antibody and E1-3-G-ABEI conjugate used as a tracer. As the results, the titer of 2E92C10 antibody was 5g/ml in soild phase and 1:2000 in liquid phase. The sensitivity on soild and solid phase CIA were about 200 pg/ml. These results indicate that CIA for measurement of E1-3-S was successfully developed by using ant-E1-3-G McAb cross-reacted with E1-3-S and could be usefully used to research this area.

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Synthesis of molecularly imprinted polymer (MIP) by radiation-induced polymerization and separation of ferulic acid from rice oil using MIP-packed column

  • Yoon, Seok-Kee;Lee, Jae-Chan;Lee, Seung-Ho;Choi, Seong-Ho;Kim, Hwa-Jung;Park, Hae-Jun;Kang, Hee-Dong
    • Analytical Science and Technology
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    • v.19 no.3
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    • pp.218-225
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    • 2006
  • A molecularly imprinted polymer (MIP) was synthesized by radiation-induced polymerization (RIP), where the ferulic acid was used as a template molecule, 4-vinylpyridine as a monomer and ethylene glycoldimethacrylate (EGDMA) as a cross-linking monomer. The MIP was packed in a glass column using a slurry method for use in medium pressure liquid chromatography (MPLC). The MPLC column was tested for separation and purification of ferulic acid from the rice oil. When repeated three times, the MPLC separation/purification yielded the ferulic acid with the purity higher than ~99%. The chemiluminescence of the luminal (5-amino-2,3-dihydro-1,4-phtalazinedione) measured on a potato disc slide (5.0 mm thick) was enhanced in the presence of ferulic acid, while, without the ferulic acid, the chemiluminescence of luminol on the potato slice disc was not observed, which suggests the ferulic acid obtained from the rice oil can be useful for immunoassay.

Prevalence of Anti-HCV among the Health-checkup Adults in Jeonbuk Province (전북 지역 건강 검진자들의 Anti-HCV 양성률 조사)

  • Kim, Yoohyun
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.1
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    • pp.32-37
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    • 2010
  • The author was performed to investigation of current status of prevalence for anti-hepatitis C virus (HCV) among the health-checkup adults in Jeonbuk province. A toal of 1,553 (male 1,046, female 507) serum samples were diagnosed by 3rd generation enzyme immunoassay (EIA) for anti-HCV. Total prevalence of anti-HCV was 0.9%, and prevalence of male and female were 0.8% and 1.2%, respectively. The prevalence of female was higher than male. According to ages group, prevalence of anti-HCV was highest in 60 age group, but it was not found in 20 age group. 14 samples with anti-HCV positive were diagnosed by EIA for hepatitis B virus surface antigen (HBs Ag), by chemiluminescence immunoassay (CLIA) for serum albumin, alanine transaminase (ALT) and asparagine transaminase (AST). Positive for HBs Ag was not found. The mean of serum albumin levels was 4.5 g/dL, and mean of ALT and AST were 34.3 IU and 31.9 IU, respectively. Through this study, I know that the prevalence of anti-HCV among adults in Jeonbuk, and suggest that the positive of anti-HCV persons who have lower serum albumin, normal to mild elevations in serum enzymes are chronic hepatitis.

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Comparison of chemiluminescence immunoassay (CIA) and immunochromatography assay(ICA) for Detecting HBsAg and Anti-HBs (B형 간염항원 및 항체 검사를 위한 화학발광면역검사법(CIA)과 면역크로마토그래피법(ICA)의 비교분석)

  • Kim, Hye-Jeong;Kim, Eun-Hee
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.7
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    • pp.3419-3424
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    • 2013
  • Diverse immunoassays including a chemiluminescent immunoassay (CIA) are used to detect hepatitis B surface antigen (HBsAg) and antibody (anti-HBs). Recently, an increasing number of institutions have been utilizing an immunochromatography assay (ICA), which is easy to use. In this study, We evaluated ICA kits for the rapid detection of HBsAg and anti-HBs by comparing them with a CIA. A total of 120 serum hospital samples, were collected for the whole month, were assayed using ICA kit. The Concordance rate, sensitivity, specificity, positive predictive value and negative predictive value of the ICA for HBsAg based on CIA results were 97%, 97%, 100%, 100%, and 96.8%, respectively. The diagnostic performances of the ICA for Anti-HBs were 90%, 90%, 93.3%, 93.1%, and 90.3%, respectively. The ICA kit failed to detect HBsAg and anti-HBs in low reactive samples. The ICA kits for the rapid detection of HBsAg might be recommended for interpreted with caution and dual analysis in the clinical laboratory.

Application of Chemiluminescence Enzyme Immunoassay Method to Collect in vivo Matured Oocyte in Dog Cloning (개 복제 시 체내 성숙 난자 회수를 위한 화학발광효소면역분석기법의 적용)

  • Kim, Min-Jung;Oh, Hyun-Ju;Kim, Geon-A;Jo, Young-Kwang;Choi, Jin;Lee, Byeong-Chun
    • Journal of Veterinary Clinics
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    • v.31 no.4
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    • pp.267-271
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    • 2014
  • Accurate determination of in vivo oocyte maturation is particularly critical for dog cloning compared to other assisted reproductive technologies because oocytes in metaphase II stage have to be recovered in order to undergo somatic cell nuclear transfer right after its recovery. The aim of present study was to evaluate the reliability and to set a reference range of a chemiluminescence enzyme immunoassay (CLEIA) compared to radioimmunoassay (RIA) method to retrieve in vivo matured oocytes. Serum progesterone concentration during proestrus and estrus was analyzed by RIA and CLEIA to determine ovulation day (Day 0). On Day 3, in vivo oocytes were recovered surgically and evaluated microscopically maturation status after staining nucleus with bisbenzimidazole dye. Mean progesterone concentration by CLEIA ($7.64{\pm}0.06ng/ml$) was significantly higher than by RIA ($6.46{\pm}0.04ng/ml$, P < 0.0001). It was not different between CLEIA ($10.01{\pm}0.34ng/ml$) and RIA values ($7.91{\pm}0.14ng/ml$, P < 0.05) on Day 0, but significantly higher CLEIA level on Day -1 and Day 1 ($6.41{\pm}0.15$ and $14.25{\pm}0.44ng/ml$) was assessed compared to RIA ($4.95{\pm}0.10$ and $11.29{\pm}0.34ng/ml$). However, with both methods, progesterone level was significantly increased from Day -1 to Day 2. To determine oocyte maturation with CLEIA method, a wider and higher reference range has to be considered.

Monitoring Ovarian Function by Solid- Phase Chemiluminescence Immunoassay

  • Kim, J.B.;Ku, P.S.
    • Clinical and Experimental Reproductive Medicine
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    • v.9 no.1_2
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    • pp.43-53
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    • 1982
  • 여성의 난소기능은 뇨중 Oestrone-3-glucuronide를 간편한 solid-phase 의 화학발광성 면역학적 측정법 (Chemiluminescence Imrnunoassay(CIA) 에 의하여 그 기능이 탐지될 수 있다. Oestrone-3-g1ucuronyl-6-bovine serum albumine에 대한 antiserum의 IgG fraction은 polystyrene 실험관벽에 흡착시켰으며, 항원으로서는 est r one-3- gl ucuronyI-6-aminoethyl-ethyl-isoluminol 을 항원 (antigen) 에 labeI 시킨 것이다. 시험 대상물인 뇨는 매일아침뇨(early morning urine) 을 희석 (1:1000 V/V)한 후 100mcl 를 취하여 이를 각기 이중분석액으로 택하였다. 시험관 내에서 결합반응 (1 hour at $4^{\circ}C)이 일어난 후에는 시험관내의 액체를 전부 흡입 폐기시켰으며, 항체반응이 일어난 후 ( antibody-bound fraction )에는 완충액 (400mcl)으로 한번 세척시켰다. 그후 염화수산화물(2N , 200mcl)을 가지고 $22^{\circ}C$에 60 분간 방치 혼합케 한 후 효소(microperoxidase) 와 과산화수소를 가하면서 산화작용에서 발생되는 발광양을 10초동안 측정하여 그 결과를 분석하였다. 위에 기술한 분석방법을 평가하면 다음과 같은 결론을 얻었다. Calibration curve sensitivity$3.12{\pm}0.75$ PG/tube ($mean{\pm}SD$)였고, lntra-assay precision(CV%) 9.52 (20 replicates;$38.4{\pm}3.66$nmol/1) 와 8.81 (15 replicates; $102.4{\pm}8.82$nmol/1)였다. Inter-assay precision(CV%) 은 11.9 (mean of 4 pools-7.03, 23.16, 52.11 과 117.53 nmol/1)로 2개월 동안에 걸쳐 시행되었고, 평균 비이어스(mean bias)는 -0.78 로 28에서 448 nmol 범위로서 매일아침 "뇨"의 차이분(different aliquots)은 좋은 결과를 얻었다. 건강한 여성으로부터 채취된 뇨중 Oestrone-3-glucuronide 의 농도(nmol/1)를 보면 월경주기의 여포기와 배난기 및 황체기에 있어서 각기 $40.2{\pm}9.9$ , $102.3{\pm}39.4$$84.3{\pm}13.3$nmol/1였다. 이와같은 결과는 동일한 검사뇨를 방사면역학적 방법(RIA)으로 측정 (6 menstrual cycle)한 결과와 유사한 측정치를 얻으므로서 간편하고 진보된 좋은 방법중의 하나라고 사료되는바이다.

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Analysis of Ochratoxin A from Deonjang, Kanjang, Gochujang Collected from Houses and Traditional Markets (가정과 재래시장에서 수거한 된장, 간장, 고추장에 존재하는 Ochratoxin A 분석)

  • 김종배;김철재;박경란;신현길
    • Journal of Food Hygiene and Safety
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    • v.9 no.4
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    • pp.221-228
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    • 1994
  • The quantitative detection of ochratoxin A (OT-A) in the traditional fermented foods were investigated to develop the analytical procedures, Enzyme-Linked Immunosorbent Assay(ELISA) and Chemiluminescence Immunoassay(CIA). Products used were divided into two groups: the first was the home-made 13 Doenjang, 12 Kanjang, and 14 Gochujang; and the second the traditional commercial products, 17 Deonjang and 11 Kanjang, which collected throughout the country. The standard curve for the quantitative determination of OT-A showed that the sensitivities in ELISA and CIA were upto the level of 20 pg/assay, and that the OT-A recovery rates were appeared to be more than 90%. The residual OT-A in the home-made products were 7.1$\pm$3.7 ng/g for Deonjang, 2.1$\pm$4.1 ng/g Kanjang were found in the traditional commercial products. Residual OT-A in the home-made products was comparatively far less than that of the traditional commercial products. At heat stability test of OT-A in the traditional fermented foods was found to be stable even at 121$^{\circ}C$ for 120 min.

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