类酶联免疫分析法可行性的研究

Study on the feasibility of enzymelinked immunosorbent assay

多环芳烃作为持久性高毒性的有机污染物,一直受到广泛关注。酶联免疫分析技术因特异性强、快速、便捷等优点而广泛使用。建立了芳香烃受体介导的类酶联免疫竞争方法(类ELISA),用于检测环境中多环芳烃类物质的含量。在实验室条件下,分别检测了5种多环芳烃萘、苊、菲、荧蒽和芘,同时比较与高效液相色谱法(HPLC)和气相色谱-质谱联用法(GC/MS)在灵敏度、准确性和重现性等标的差异。HPLC检出限为2.70~10.71 ng/mL;GC/MS的检出限为2.60~24.40 ng/mL;类ELISA的检出限在1.24~3.10ng/mL。HPLC的加标回收率为58.03%~106.69%;GC/MS的加标回收率为88.16%~154.83%;类ELISA的加标回收率为74.67%~121.84%。HPLC的变异系数为4.26~9.03;GC/MS的变异系数为1.44~5.82;类ELISA的变异系数为2.56~8.16。结果表明,类ELISA方法灵敏度较好,准确性较高;而GC/MS方法重现性较好,检测更为稳定。

The present paper is aimed at introducing the experimental result by establishing an aromatic hydrocarbon receptormediated enzyme-linked immunoassay( ELISA) through detecting the content of the polycyclic aromatic hydrocarbons in the environment. For this purpose,we have adopted the enzymelinked immunoassay technology through the analytical test. As a matter of fact,the above said ELISA method enjoys a strong specificity and sensitivity for its great convenience as compared with the traditional analytical methods. Furthermore,we have also tested 5 polycyclic aromatic hydrocarbons under the laboratory conditions, naphthalene, anthracene, phenanthrene, fluoranthene and anthracene,to identify and determine the feasibility of the established method and gain the key testing parameters,such as the sensitivity,accuracy and reproducibility. Besides,we have also calculated and worked out the detection limits,the spike recovery rate,and the variation coefficient. Just under the same conditions through the comparison with the high performance liquid chromatography( HPLC) and the gas chromatography-mass spectrometry( GC/MS) that we have found and determined that the experimental result has turned out quite in contrast. More clearly speaking,the detection limit of HPLC can be found ranging from 2. 70 to 10. 71 ng/mL,whereas the detection limit of GC/MS may range from 2. 60 to 24. 40 ng/mL and the detection limit of ELISA has been in a range of 1. 24-3. 10 ng/mL. At the same time,the recovery range of HPLC can also be found and determined by the range from 58. 03% to106. 69%,while the recovery range of GC/MS should be stated between 88. 16% and 154. 83% and the spiked recovery range of ELISA stands from 74. 67% to 121. 84%. Thus,correspondingly,the coefficient variation of HPLC has to be 4. 26-9. 03,while,in contrast,the coefficient of the variation of GC/MS should be between 1. 44 to 5. 82,with the coefficient of variation of ELISA being 2. 56-8. 16. Hence,it can be seen from the above data that the detection limit of the ELISA-like method should be the lowest. On the other hand,the standardized recovery rate should be more appropriate to meet the standardized need than the other related methods,with the coefficient of variation also coming to conform to the standard of the analytical method. Moreover,it can also be detected that the analytical method ought to be feasible to detect the polycyclic aromatic hydrocarbons in the environment by the enzyme-linked immunosorbent assay. Besides,the ELISA-like method can be made more sensitive and accurate,too,by the enzyme-linked immunosorbent assay. For instance,when the number of samples is great enough,it should be possible to choose the enzyme-linked immunoassay for screening. The reason for this is that the GC/MS method enjoys the lowest variation coefficient to work out better reproducibility and stable detection for the effective quantitative testing.