|本期目录/Table of Contents|

[1]刘 冉,张世联,牟振云,等.牛乳中抗幽门螺杆菌特异性抗体的 间接ELISA法的建立[J].医学研究与战创伤救治(原医学研究生学报),2008,10(03):174.
　LIU Ran,ZHANG Shi-lian,MOU Zhen-yun,et al.Development of indirect enzyme-linked immunosorbent assay(ELISA) for detection of anti-Helicobacter pylori antibodies from bovine immune milk[J].JOURNAL OF MEDICALRESEARCH —COMBAT TRAUMA CARE,2008,10(03):174.
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牛乳中抗幽门螺杆菌特异性抗体的 间接ELISA法的建立()
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《医学研究与战创伤救治》（原医学研究生学报）[ISSN:1672-271X/CN:32-1713/R]

卷:
第10卷
期数:
2008年03期
页码:
174
栏目:
出版日期:
2008-05-20

文章信息/Info

Title:
Development of indirect enzyme-linked immunosorbent assay(ELISA) for detection of anti-Helicobacter pylori antibodies from bovine immune milk
文章编号:
1672-271X(2008)03-0174-04
作者:
刘 冉1; 张世联2;  牟振云3;  王 薇2
1.东南大学公共卫生学院, 江苏南京 210009;2. 河北省疾病预防控制中心, 河北石家庄 050021; 3. 河北医科大学, 河北石家庄 050020
Author(s):
LIU Ran1; ZHANG Shi-lian2; MOU Zhen-yun3; WANG Wei2
1.School of Public Health,Southeast University,Nanjing，210009,Jiangsu,China; 2. Hebei Center for Disease Control and Prevention,Shijiazhuang 050021,Hebei,China; 3. Hebei Medical University, Shijiazhuang 050020,Hebei,China
关键词:
幽门螺杆菌; 免疫乳; 间接ELISA
Keywords:
Helicobacter pylori;  Immune milk;  Indirect-ELISA
分类号:
R392.11；R573
DOI:
-
文献标志码:
A
摘要:
目的 建立抗幽门螺杆菌(Hp)特异性抗体的ELISA检测方法，用于牛乳中抗Hp特异性抗体的检测。方法 超声Hp全菌抗原接种奶牛，按多克隆抗体制备技术制备牛抗Hp抗血清。用纯化牛IgG免疫家兔，制备兔抗牛IgG多克隆抗体。硫酸铵盐析，DEAE-32柱层析分别纯化牛与兔IgG。兔抗牛IgG以辣根过氧化物酶标记。用Hp全菌抗原包被反应板，建立间接ELISA法用于牛乳抗Hp抗体的检测。 结果 建立的抗Hp间接ELISA法的最佳包被抗原量为1.47μg/孔，HRP-兔抗牛IgG 1∶600稀释，标准曲线方程为A=0.0124+0.3988 lnC，相关系数r=0.999 8，线性范围为1.8～145 μg/ml,回收率在86.4％～92.8％，变异系数为9.4％～12.3％。结论 初步建立牛乳中抗Hp抗体的间接ELISA测定法，可用于Hp免疫牛乳中抗Hp特异性IgG的检测。
Abstract:
Objective To develop an ELISA method for specific detection of anti-Helicobacter pylori antibodies from bovine immune milk. Methods Cows were immunized routinely with Hp sonicate antigens, and rabbits were immunized with purified bovine IgG. Bovine anti-Hp and rabbit anti-bovine polyclonal antibodies were purified with ammonium sulfate and DEAE-32. Indirect-ELISA was developed for detection of anti-Helicobacter pylori IgGs in milk with HRP-rabbit anti-bovine IgGs. Results The optimal working dose of coating antigens was 1.47 μg/well and the dilution of HRP-rabbit anti-bovine IgGs was 1∶600. The standard equation of the indirect-ELISA was OD=0.0124+0.3988 LnC and the correlation coefficient was 0.999 8. Linear range of anti-Hp IgG detection was 1.8~145 μg/ml. The anti-Hp IgG recovery rate varied from 86.4% to 92.8% with the range of coefficient of variation as 9.4%~12.3%. Conclusion The indirect-ELISA developed in this study can be used to detect anti-Hp specific IgG in bovine immune milk.

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备注/Memo

备注/Memo:
河北省医学科研基金项目(01118)
更新日期/Last Update: 2013-11-22