不同品牌ELISA试剂盒检测细胞培养上清液中IL-8对比研究

Comparative study on IL-8 detection in cell culture supernatant by ELISA kit of different brands

目的 对比研究2种不同品牌ELISA试剂盒检测细胞培养上清液中人白细胞介素-8(IL-8)水平情况。方法 不同浓度聚肌胞苷酸(Poly I:C)刺激A549细胞10、24 h,收集细胞上清液,共获得40份样本,分为1~10组,每组4份。采用2种ELISA试剂盒(试剂盒A、试剂盒B)检测细胞培养上清液中IL-8水平,对比2个试剂盒检测得到的组间数据或同组数据的一致性或差异性。结果 2种试剂盒拟合曲线中相关系数值分别为:0.999 600、0.999 602,检测标准品结果相关性良好。试剂盒B检测样品1、2组中IL-8水平比较,差异有统计学意义(P<0.01),但试剂盒A检测样品1、2组中IL-8水平比较,差异无统计学意义(P>0.05);2种试剂盒检测样品8、9组及样品9、10组中IL-8水平比较,差异均有统计学意义(P<0.01)。仅试剂盒B检测样品1、5组中IL-8水平明显低于试剂盒A,差异有统计学意义(P<0.01)。结论 不同品牌ELISA试剂盒均可用于检测IL-8水平,但检测结果仍在一定程度上存在差异。为避免实验误差,研究中应尽量选择同一个厂家生产的同品牌试剂盒。

Objective To compare the difference of human interleukin-8(IL-8)in cell culture supernatant detected by two commercial ELISAs.Methods After stimulating A549 cells with different concentrations of polyinosine(PolyI:C)for 10 and 24 hours,the cell supernatant was collected,and a total of 40 samples were obtained,which were divided into 1 to 10 groups with four samples per group.Two ELISA kits(Kit A,kit B)were used to detect IL-8 levels in the supernatant of cell culture,and the consistency or difference of the data detected by the two kits between groups or within the same group was compared.Results The correlation coefficient values in the fitting curves of the two kits were 0.999600 and 0.999602,respectively,indicating a good correlation between the results of test standards.There was statistical significance in IL-8 level between group 1 and 2 of samples detected by kit B(P<0.01),but there was no statistical significance in IL-8 level between group 1 and 2 of samples detected by kit A(P>0.05).There were significant differences in IL-8 levels between groups 8 and 9 of samples and groups 9 and 10 of samples detected by the two kits(P<0.01).The level of IL-8 in groups 1 and 5 of samples detected by kit B was significantly lower than that of kit A,and the difference was statistically significant(P<0.01).Conclusion Different brands of ELISA kits can be used to detect IL-8 levels,but the detection results are still different to a certain extent.In order to avoid experimental errors,the same brand kit produced by the same manufacturer should be selected as far as possible in the study.