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目的建立石头鱼毒素neoVTX的酶联免疫吸附试验(ELISA)检测方法,为该毒素的检测、中毒诊断及治疗提供依据。方法以过碘酸钠法偶联马抗-HRP,利用双抗体夹心ELISA方法检测neoVTX。结果该法的线性范围在磷酸盐缓冲液(PBS)中为4~512 ng/ml,线性回归方程为Y=1.676X-1.140(r~2=0.9728,n=8,P<0.0001),在血浆中检测线性范围为4~2048 ng/ml,线性回归方程为Y=0.3094X-0.1208(r~2=0.9907,n=10,P<0.0001)。牛血清白蛋白、卵清蛋白及人血浆对检测结果无干扰。结论成功建立了石头鱼毒素的双抗体夹心ELISA检测方法,该法有较高的灵敏度和特异性,适用于溶液中及血浆中微量neoVTX的样品分析。
OBJECTIVE To establish an enzyme-linked immunosorbent assay (ELISA) for the detection of neotoxin, and to provide a basis for the detection, diagnosis and treatment of the toxin. Methods The anti-HRP was coupled by sodium periodate and the neoVTX was detected by double antibody sandwich ELISA. Results The linear range of this method was 4 ~ 512 ng / ml in phosphate buffered saline (PBS). The linear regression equation was Y = 1.676X-1.140 (r ~ 2 = 0.9728, n = 8, P <0.0001) The linear range of plasma detection was 4-2048 ng / ml. The linear regression equation was Y = 0.3094X-0.1208 (r = 2 = 0.9907, n = 10, P <0.0001). Bovine serum albumin, ovalbumin and human plasma on the test results without interference. Conclusion The double antibody sandwich ELISA assay was successfully established. It has high sensitivity and specificity and is suitable for sample analysis of micro neoVTX in solution and in plasma.