Development of a supersensitive detection method for pancreas cancer associated antigen and tumor suppressor gene by using immuno-PCR.
利用免疫PCR开发胰腺癌相关抗原和抑癌基因的超灵敏检测方法。
基本信息
- 批准号:07557046
- 负责人:
- 金额:$ 3.9万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (B)
- 财政年份:1995
- 资助国家:日本
- 起止时间:1995 至 1996
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
A sensitive method for the detection of antigens in sera, termed double determinant immuno-polymerase chain reaction (DDI-PCR), was developed using two monoclonal antibodies (MoAbs) in which the antigens are sandwiched and a specific DNA molecule as a marker. Instead of the antigen itself, the first MoAb to bind the circulating antigens was immobilized. After the biotinylated second MoAb was bound to the antigen, free streptavidin was used to attach a biotinylated DNA to the biotinylated second MoAb. The biotinylated DNA complexed with antigen-antibody-streptavidin was amplified by PCR.The PCR products were analyzed by Southern blot hybridization after agarose gel electrophoresis. Compared with the conventional ELISA using soluble intercellular adhesion molecule-1 (sICAM-1) in the supernatant of cultured Panc-1 cells as an antigen, our DDI-PCR was 10^3 times more sensitive in detection limit. In both the culture medium and sera from gastric cancer patients of high sICAM-1 titer, an approximately 10^3 -fold enhancement in detection sensitivity was obtained compared with ELISA.In addition, the DDI-PCR system can detect the antigen in sera at a level below the detection limit of traditional ELISA methods with high sensitivity. Thus, DDI-PCR has the significant advantage that it can be readily applied to any antigen-antibody system with two MoAbs without making any original molecules.
利用双抗体夹心法和特异性DNA分子作为标记物,建立了一种灵敏的检测血清中抗原的方法,即双决定簇免疫聚合酶链反应(DDI-PCR)。固定化的不是抗原本身,而是结合循环抗原的第一个单克隆抗体。在生物素化的第二MoAb与抗原结合后,使用游离的链霉亲和素将生物素化的DNA连接到生物素化的第二MoAb。将生物素标记的DNA与抗原-抗体-链霉亲和素复合后进行PCR扩增,PCR产物经琼脂糖凝胶电泳后进行Southern印迹杂交分析。与以Panc-1细胞培养上清中可溶性细胞间粘附分子-1(sICAM-1)为抗原的常规ELISA法相比,该方法的灵敏度提高了103倍。对sICAM-1高滴度的胃癌患者血清和培养液的检测灵敏度均比ELISA法提高约103倍,且检测灵敏度高,可在低于传统ELISA法检测限的水平上检测血清中的抗原。因此,DDI-PCR具有显著的优点,即它可以容易地应用于具有两个MoAb的任何抗原-抗体系统,而无需制备任何原始分子。
项目成果
期刊论文数量(7)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Itoh, F.et al.: "Double determinant immuno-polymerase chain reaction for detecting sICAM-1." Artificial Organs. 20 (8). 898-901 (1996)
Itoh, F.et al.:“用于检测 sICAM-1 的双决定簇免疫聚合酶链反应。”
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
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- 通讯作者:
Suzuki, A.et al.: "Double determinant immuno-polymerase chain reaction : A sensitive method for detecting circulating antigen in human sera." Jpn. J.Cancer Res.86. 885-889 (1995)
Suzuki, A.等人:“双决定簇免疫聚合酶链反应:一种检测人血清中循环抗原的灵敏方法。”
- DOI:
- 发表时间:
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- 影响因子:0
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- 通讯作者:
Shijubo, N.et al.: "Circulating soluble intercellular adhesion molecule-1 (sICAM-1) in patients with sarcoidosis." Clin. Exp.Immunol.106. 549-554 (1996)
Shijubo, N.等人:“结节病患者的循环可溶性细胞间粘附分子-1 (sICAM-1)。”
- DOI:
- 发表时间:
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- 影响因子:0
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Makiguchi, Y.et al.: "Effect of MUC1 mucin, an anti-adhesion molecule, on tumor cell growth." Jpn. J.Cancer Res.87. 505-511 (1996)
Makiguchi, Y.等人:“MUC1 粘蛋白(一种抗粘附分子)对肿瘤细胞生长的影响。”
- DOI:
- 发表时间:
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- 影响因子:0
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- 通讯作者:
Makiguchi,Y.,et al: "Effect of MUC1 mucin,an anti-adhesion molecule,on tumor cell growth." Jpn.J.Cancer Res.87. 505-511 (1997)
Makiguchi, Y. 等人:“MUC1 粘蛋白(一种抗粘附分子)对肿瘤细胞生长的影响。”
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IMAI Kohzoh其他文献
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{{ truncateString('IMAI Kohzoh', 18)}}的其他基金
Identification of specific cancer stem cell markers for diagnosis
鉴定用于诊断的特定癌症干细胞标记物
- 批准号:
24650632 - 财政年份:2012
- 资助金额:
$ 3.9万 - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
Development of novel gastrointestinal cancer diagnostic system based on large epige-nome analysis.
基于大表观基因组分析的新型胃肠道癌症诊断系统的开发。
- 批准号:
23240129 - 财政年份:2011
- 资助金额:
$ 3.9万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
Scientific Support Programs for Cancer Research Grant-in-Aid for Scientific Research on Innovative Areas
癌症研究科学支持计划 创新领域科学研究资助计划
- 批准号:
221S0001 - 财政年份:2010
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$ 3.9万 - 项目类别:
Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
Molecular mechanisms of epigenetic alterations in gastrointestinal cancer and application to diagnosis and therapy
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- 批准号:
17109008 - 财政年份:2005
- 资助金额:
$ 3.9万 - 项目类别:
Grant-in-Aid for Scientific Research (S)
Immunotherapy using humanized monoclonal antibody
使用人源化单克隆抗体的免疫疗法
- 批准号:
17016060 - 财政年份:2005
- 资助金额:
$ 3.9万 - 项目类别:
Grant-in-Aid for Scientific Research on Priority Areas
Microsatellite instability and aberrant methylation in gastrointestinal cancer and its application to molecular target therapy
胃肠道肿瘤微卫星不稳定性和异常甲基化及其在分子靶向治疗中的应用
- 批准号:
13854016 - 财政年份:2001
- 资助金额:
$ 3.9万 - 项目类别:
Grant-in-Aid for Scientific Research (S)
Immunelogical diagnosis of MSI-positive gastrointestinal cancer and application for cancer vaccine
MSI阳性胃肠癌的免疫学诊断及癌症疫苗的应用
- 批准号:
12470124 - 财政年份:2000
- 资助金额:
$ 3.9万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Analysis of molecular mechanism of premalignant lesion of the stomach using genomic instability and new mucin gene.
利用基因组不稳定性和新粘蛋白基因分析胃癌前病变的分子机制。
- 批准号:
11557043 - 财政年份:2000
- 资助金额:
$ 3.9万 - 项目类别:
Grant-in-Aid for Scientific Research (B).
AnalysIs of molecular mechanism of gastric pre-cancerous lesions by using new mucin genes
新粘蛋白基因分析胃癌前病变的分子机制
- 批准号:
09470141 - 财政年份:1997
- 资助金额:
$ 3.9万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Detection of Novel Amyloid-Associated Protein by Monoclonal Antibody and its Gene Analysis
新型淀粉样蛋白相关蛋白的单克隆抗体检测及其基因分析
- 批准号:
01570363 - 财政年份:1989
- 资助金额:
$ 3.9万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
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