主管：中华人民共和国司法部
主办：司法鉴定科学研究院
ISSN 1671-2072  CN 31-1863/N
中国司法鉴定

中国司法鉴定 ›› 2012 ›› Issue (1): 16-19.

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定量检测毒品的蛋白芯片的研发

曾立波， 陈连康， 胡小龙等   

  1. 上海市公安局物证鉴定中心，上海市现场物证重点实验室，上海 200083
  • 收稿日期:2011-11-10 出版日期:2012-01-15 发布日期:2022-07-12
  • 作者简介:曾立波（1954-），男，主任法医师，主要从事分子免疫学研究和法医毒物、毒品分析。
  • 基金资助:
    “十一五”国家科技支撑计划项目（2006038022001）

Development of Protein Chips for Quantitative Drugs Assay

ZENG Li-bo， CHEN Lian-kang， HU Xiao-long, etc.   

  1. Shanghai Key Laboratory of Crime Scene Evidence， Institute of Forensic Science， Shanghai Municipal Public Security Bureau， Shanghai 200083， China
  • Received:2011-11-10 Published:2012-01-15 Online:2022-07-12

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摘要：

目的 利用蛋白芯片技术建立一种快速、高通量的毒品定量检测方法。方法 采用硝酸纤维膜作为蛋白芯片的基片，将多种毒品和蛋白偶联物抗原包被在芯片基片上后封闭，不同毒品的单克隆抗体与之结合，再用荧光染料Cy5标记的二抗孵育，样本中如果含有毒品成分，将会与包被的毒品偶联物竞争性地与毒品的抗体相结合，导致最终的荧光信号发生改变，荧光信号的变化与样品中的毒品的浓度相关。通过荧光芯片检测仪CCD可以进行定量分析，最终判断样本中毒品的含量。结果 对506例样本进行定性定量检测，与GC-MS仪器比较后总相关性在88%以上，芯片的检测灵敏度比胶体金方法提高10倍，其检测的特异性达到99%。结论 蛋白芯片是一种有效、准确的高通量检测毒品含量的方法。

关键词: 毒品毒物分析, 吗啡, 甲基苯丙胺, 蛋白芯片, 免疫检测

Abstract:

Objective To develop a high-throughout quantitative assay method for detecting drugs in human body fluid samples. Methods By using the nitrocellulose membrane as the matrix of protein chips， the membrane was treated to generate active groups for protein binding. After dispersing the conjugates of the drug molecules with the carrier protein， the membrane was blocked. Different antibodies for drugs were labeled with fluorescent dye CY5， which would generate fluorescent signal when combining to the conjugates. If there are target drugs in the sample， the drugs will competitively combine to the conjugates and the fluorescent signal will decrease. The fluorescent readers can analyze the intensity of the signal and quantitatively assay the concentration of the drugs in the sample. Results The results of totally 506 samples tested by the protein arrays for drugs had a correlation of more than 88% with the GC/MS assay. The results showed that it can specifically detect drugs such as morphine， methamphetamine， and so on， without any cross reaction. The sensitivity of the assay is 10ng/ml， which is 10 times higher than that of the gold colloid test. Conclusion The protein chips is an effective， accurate and high-throughout assay for detecting drugs in human body fluid samples.

Key words: forensic toxicological analysis, morphine, methamphetamine, protein chip, immunoassay

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