食品科学 ›› 2009, Vol. 30 ›› Issue (18 ): 274-277.doi: 10.7506/spkx1002-6630-200918061

• 分析检测 • 上一篇    下一篇

呋喃西林代谢物氨基脲免疫吸附生物条码检测方法的初步研究

唐 勇,王五洲,向军俭* ,王 宏,邓 宁,杨红宇   

  1. 暨南大学抗体工程研究中心
  • 收稿日期:2009-07-03 出版日期:2009-09-15 发布日期:2010-12-29
  • 通讯作者: 向军俭* E-mail:txjj@jnu.edu.cn
  • 基金资助:

    广东省自然科学基金项目(7300094)

Primary Study on Immunosorbent Bio-Barcode Assay for the Determination of Semicarbazide, A Metabolite of Furacilin

TANG Yong,WANG Wu-zhou,XIANG Jun-jian*,WANG Hong,DENG Ning,YANG Hong-yu   

  1. Center of Antibody Engineering, Jinan University, Guangzhou 510632, China
  • Received:2009-07-03 Online:2009-09-15 Published:2010-12-29
  • Contact: XIANG Jun-jian*, E-mail:txjj@jnu.edu.cn

摘要:

制备复合物抗氨基脲衍生物(CPSEM)单抗- 胶体金- 单链硫醇化DNA(anti-CPSEM-Au-ssSHDNA),建立氨基脲(SEM)的新型高灵敏度的免疫吸附生物条码检测方法。制备腹水型抗CPSEM 单抗,饱和硫酸铵法纯化,然后将其偶联到柠檬酸三钠还原法制备的20nm 胶体金纳米颗粒表面,制备二者的偶联物,再在单抗- 纳米金偶联产物的基础上连接硫醇化DNA,结合竞争ELISA 原理和PCR 技术将ELISA 中的酶信号转化为可扩增的DNA 信号,建立新型的免疫吸附生物条码检测方法。本研究制备出抗CPSEM 单抗- 纳米金- 单链硫醇化DNA 复合物,初步建立的检测SEM的免疫吸附生物条码检测法灵敏度为8pg/ml。

关键词: 呋喃西林, 氨基脲, 胶体金, 单链硫醇化DNA

Abstract:

Abstract:This study aimed to develop a new and sensitive immunosorbent biobarcode assay the detection of semicarbazide (SEM), a metabolite of nitrofurazone based on the conjugation of Anti-CPSEM-Au-ssSHDNA. Ascetic monoclonal antibody against CPSEM purified by ammonium sulfate precipitation and ssSHDNA were conjugated to Au nanoparticles with a diameter of 20 nm prepared by controlled reduction of gold chloride with sodium citrate. Then a novel immunosorbent bio-barcode assay, an integration of PCR and idc-ELISA, which can transform enzyme signal into DNA signal, could be developed based on the conjugation of Anti-CPSEM-Au-ssSHDNA. An anti-CPSEM-Au-ssSHDNA complex was successfully prepared, and the immunosorbent bio-barcode DNA detection assay, which could detect 8 pg /ml semicarbazide, was primarily developed.

Key words: furacilin, semicarbazide (SEM), colloidal gold, ssSHDNA

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