免疫胶体金法提取环境标本中细菌DNA技术

将抗-DNA单克隆抗体标记在胶体金颗粒上制成免疫胶体金试剂，提取标本中DNA，直接用于PCR检测，从而建立一种简单、快速、高效的免疫胶体金方法提取环境标本中的DNA。结果表明：应用免疫胶体金试剂可有效去除环境标本中PCR抑制剂，浓缩模板，提高PCR检测敏感度3～4个数量级。操作步骤简单，无需使用有机溶剂，避免环境污染，吸附了DNA的免疫胶体金可直接用于PCR扩增。研制了免疫胶体金试剂并确定其最佳反应条件，有效提高PCR技术在检测现场环境标本中的敏感性和实用性。

Extraction of DNA Using Monoclonal Anti-DNA and Colloidal-gold Beads

To developing a simple, rapid and sensitive immunoaffinity method for extracting DNA before amplification by the polymerase chain reaction(PCR). Using monoclonal anti-DNA bound to colloidal-gold beads(which called immunocolloidal-gold ) and extracting DNA from the specimen before amplification by PCR. The immunoaffinity method we use is extremely efficient in extracting DNA at low concentration, also, the beads carrying the DNA/anti-DNA complexes can be added directly to a PCR mixture without elution, and the presence of PCR inhibitors in specimens has no effect on amplification. When applied to serial dilution bacteria suspension, the detection of limit can be 100 CFU/mL, and the sensitivity of detecting artificial soil sample and milk sample were 101 and 100 CFU/mL. The immunoaffinity method described here can sever as a sensitive and practicable method for extracting DNA , particularly where samples have low concentrations of DNA or where the material is in poor condition.