| 摘要: | 甲基安非他命 ( methamphetamine ) 最初用來治療像是精神分裂症中樞神經系統失調的疾病，在世界各地，這類神經刺激物質變成主要的濫用藥物之一。高劑量使用甲基安非他命會引發活動過度、不安、發燒、高血壓、心毒性和類似妄想症的神經失調疾病。本研究的目的主要是甲基安非他命抗體之生產，並應用此抗體於免疫層析試片 ( immunochromatographic strip ) 之開發。本實驗工作第一部分成它X成出藥物與蛋白質之生物共軛物，其中甲基安非他命與牛血清蛋白之生物共軛物 ( Meth-BSA ) 當作免疫原 ( immunogen ) 進行小鼠免疫工作；另外，甲基安非他命與絲膠蛋白之生物共軛物 ( Meth-Ser ) 當作 ELISA 檢測系統之抗原 ( antigen ) 。第二部分生產並純化出 5.68 mg/mL甲基安非他命多株抗體 ( anti-Meth pAb ) 。第三部分成市堨艂K疫金層析試片系統，所需樣品量為80 μL， 能夠檢測濃度為10 ～50 mg/mL 的甲基安非他命且檢測所需時間約15-30分鐘。試片保存期限於4℃ 可達28天以上。此試片的開發可應用於咖啡、可樂、綠茶、尿液及小鼠血清之實際樣品檢測。本研究所製備之免疫層析試片，具有以下優點：1. 高度專一性；2. 靈敏度高；3. 不需前處理；4. 所需樣品量少；5. 操作方便；6. 檢測快速；7. 低成本；8. 沒有交叉反應；9. 可長期保存；10. 不需要任何儀器之使用等優點。因此本實驗所製備之免疫層析試片極具應用價值。
Methamphetamine ( Meth ) is a drug that was initially synthesized to treat central nervous system ( CNS ) disorders such as schizophrenia. This psychostimulant has now become a major drug of abuse worldwide. In high doses, methamphetamine causes hyperactivity, agitation, hyperthermia, hypertension, cardiotoxicity and psychotic disorder such as paranoia. The objective of this study was to produce polyclonal antibody against methamphetamine. The antibody will be used for the application in the development of a nano-particle colloidal gold immunochromatographic strip. In this study, firstly, the drug-proteins bioconjugates had been successfully produced. Meth-BSA bioconjugate was as the immunogen and Meth-sericin bioconjugate was as the antigen. Secondly, production and purification of polyclonal antibody against methamphetamine were made. The pAb concentration was 5.68 mg/mL. Finally, a self-assembled nano-gold immunochromatographic strip was established. The sample required was 80 μL. The concentration range for detection of Meth using this self-assembled immunochromatographic strip was 10 ～50 mg/mL and determination time was 15～30 min. The storage time could be up to 28 days at 4 ℃. The Meth immunochromatographic strip could be applied to test coffee, Coca-Cola, green tea, urine and mice serum. The advantages of the self-assembled immunochromatographic strip are high specificity, high sensitivity, no pretreatment, low sample requirement, easy operation, rapid determination, low price, no cross-reaction, long-term preservation and no machine needed etc. |
