胶体金免疫层析法同步快速测定畜禽肉中3种兽药残留

该文旨在建立一种胶体金免疫层析技术同步测定猪肉和鸡肉组织中呋喃唑酮代谢物、呋喃西林代谢物、氯霉素药物残留的方法。动物肌肉组织中呋喃唑酮代谢物、呋喃西林代谢物、氯霉素经酸化、衍生化及乙酸乙酯提取,并经含吐温-20的Tris-HCl缓冲液稀释后,与胶体金标记的单克隆抗体结合,抑制抗体和硝酸纤维素膜检测线(T线)上的抗原结合,从而导致T线颜色变化。通过比较T线和质控线(C线)颜色深浅,对试样中多兽药残留进行定性判定。试验优化了pH值、抗体添加量、抗原包被量、吐温-20添加量、衍生化试剂浓度和冻干保护剂种类等关键因素,并对检测卡的灵敏度、特异性和稳定性进行检测。结果显示,所建立的免疫层析方法可在35 min内检出猪肉和鸡肉中呋喃西林代谢物、呋喃唑酮代谢物和氯霉素药物,检出限分别为0.5、0.5、0.1μg/kg,灵敏度≥98%,假阳性率≤2%,假阴性率≤2%,稳定性可达到1年,且与现有标准方法检测结果一致。该方法操作简便、特异性强、灵敏度和准确度高,检测时间短且稳定性好,可用于动物肌肉组织中多兽药残留的高通量快速检测。

GICA检测高致病性禽流感抗体方法的建立

〔目的〕 建立可快速、简便地检测微量禽流感的抗体的方法。〔方法〕 利用灭活的高致病性禽流感病毒H5N1建立禽流感病毒浓缩纯化样品体系,获得大量纯度较高的禽流感病毒(1:5 12 )。将2 .80mg/mL纯化病毒与一定大小的胶体金颗粒偶联,吸附在玻璃纤维上,制作的高致病性禽流感病毒抗体胶体金检测试纸条,用于高致病性禽流感定量(合格)检测。〔结果〕 显色反应良好,条带稳定,检测获得成功。〔结论〕 其反应特异性、敏感性、重复性良好,具备微量、快速、简便的特点,适合于进出境现场检疫和疫情普查。

A novel colloidal gold immunochromatography assay strip for the diagnosis of schistosomiasis japonica in domestic animals

Background:Schistosomiasis remains a major public health concern in China and an epidemiological survey has revealed that schistosome-infected bovines and goats are the main transmission sources for the disease.Therefore,development of a sensitive technique for the diagnosis of schistosomiasis in domestic animals is necessary.Method:A novel colloidal gold immunochromatography assay(GICA)strip was developed for detecting Schistosoma japonicum in domestic animals.The colloidal gold was conjugated with recombinant streptococcal protein G(rSPG).As the test and control lines,the schistosome soluble egg antigen and rSPG,respectively,were blotted on nitrocellulose membrane.Results:The lowest detectable serum dilution was 1∶640 for schistosome-infected buffaloes.The cross-reaction rate of GICA was 14.29%with Paramphistomum sp.in buffaloes,16.67%with Haemonchus sp.in goats,and 33.33%with Orientobilharzia sp.in goats.These results were slightly lower and similar to those obtained through ELISA.Moreover,the strips for detecting S.japonicum in mice,rabbits,buffaloes,and goats showed high sensitivity(100.00%,100.00%,100.00%,and 100.00%,respectively)and specificity(100.00%,100.00%,94.23%,and 88.64%,respectively).And the sensitivity or specificity of the GICA strips did not present any significant differences after storage for 12 months at room temperature.When compared with ELISA,the GICA strips exhibited similar sensitivity and specificity in the diagnosis of schistosomiasis in mice,rabbits,buffaloes,and goats.Besides,only 5μl of serum are required for the test and the detection can be completed within 5 min.Conclusion:This study is the first to develop a GICA strip using gold-rSPG conjugate for the diagnosing of schistosomiasis in domestic animals,and preliminary results showed that the developed strip may be suitable for large-scale screening of schistosomiasis in endemic areas.

胶体金免疫层析法快速检测桔青霉素的研究

目的:开发一种快速、稳定的现场检测谷物中桔青霉素(Citrinin, CIT)的方法。方法:将桔青霉素与载体蛋白BSA和HSA连接制备人工抗原,将人工抗原免疫Babl/c小鼠制备特异单克隆抗体(2D3)制备胶体金-mAb探针和免疫层析试纸条。结果:单克隆抗体的亲和力为7.17×10~8 L/mol,与其它霉菌毒素无交叉反应。在最佳条件下,视觉检测限和仪器检测限分别为5 ng/mL和0.11 ng/mL。免疫层析(ICA)技术对玉米和小麦样品的添加回收率为86.93%~110.34%,相对标准偏差为0.47%~2.59%,检测结果与高效液相色谱(HPLC)法具有良好的一致性(R^2>0.98)。结论:胶体金免疫层析技术可用作现场检测CIT含量的快速、可靠的技术。

Study on Rapid Detection of Tetracyclines,Fluoroquinolones and Sulfonamides in Milk

Aiming at the market demand for rapid detection of tetracyclines,fluoroquinolones and sulfonamides in milk,a golloidal gold immunochromatography test strip for simultaneous detection of tetracyclines,fluoroquinolones and sulfonamides in milk was prepared based on the principle of competitive inhibition immunochromatography. The performance indicators of the test strip were verified. The results showed that the test strip can simultaneously detect 4 tetracyclines,13 fluoroquinolones and 13 sulfonamides,and the detection limits all can meet the national residue limits; the tests strip exhibited false positive rate≤5% and false negative rate = 0; and no cross-reaction with other drugs was commonly found in milk,indicating good specificity. The method is simple,rapid,and has low cost and easy popularization. It provides a means for realizing on-site rapid detection and is of important practical significance to guarantee of safety of milk and dairy products in China.

胶体金免疫层析法快速检测韭菜中腐霉利残留

【目的】建立韭菜中腐霉利残留的胶体金免疫层析快速检测方法。【方法】通过合成两种不同结构的腐霉利半抗原,并分别与载体蛋白耦联制得免疫原和包被原,免疫小鼠获得腐霉利单克隆抗体。基于竞争性免疫抑制原理,建立可视化胶体金免疫层析法。【结果】检测韭菜中腐霉利残留,检测限为5 mg/kg,满足国家标准规定的最大残留限量;对实际样本的检测结果与液相色谱-质谱法检测结果一致。【结论】本研究建立的方法具有简便、快速、准确等优点,可实现对韭菜中腐霉利残留超标的高通量快速筛选。