龈沟液微量样本中多种成分的检测

目的:以龈沟液(gingival crevicular fluid,GCF)样本为例,探寻微量样本中多种成分的检测方法.方法:根据酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)的基本原理,确认待检测成分之间无交叉反应,样本与包被抗体在4 ℃下进行低温反应,重复使用同一样本.39份龈沟液来自4例轻、中度牙周炎患者,龈沟液洗提液分为两份,一份样本纳入A组,用ELISA方法先检测白细胞介素6(interleukin-6,IL-6),再用此样本检测白细胞介素1β(interleukin-1β,IL-1β);另一份样本纳入B组,直接进行IL-1β检测.比较A组与B组IL-1β的检测结果.65份龈沟液样本选自9例来北京大学口腔医院修复科就诊患者,龈沟液洗提液分为两份,一份样本纳入C组,用ELISA方法先检测肿瘤坏死因子(tumor necrosis factor-α,TNF-α)后,再用此样本检测弹性蛋白酶;另一份纳入B组,直接进行弹性蛋白酶检测.比较A组与B组弹性蛋白酶的检测结果.结果:A和B组IL-1β光密度值分别为(0.5±0.4)和(0.5±0.4),差异无统计学意义(P=0.136),且高度相关(r=0.993,P=0.000);C和D组弹性蛋白酶吸光度值分别为(1.1±0.6)和(1.1±0.6),差异无统计学意义(P=0.943),且高度相关(r=0.979,P=0.000).结论:通过重复使用同一样本,可以达到用微量样本检测多种成分的目的.

Detection of various component in the trace sample of gingival crevicular fluid

OBJECTIVE: To evaluate methods of detecting various component in the trace sample of the gingival crevicular fluid (GCF). METHODS: One sample of GCF was collected from four patients with mild or moderate periodontitis. At first, one half (group A)of every sample was measured for interleukin-6 (IL-6) by enzyme-linked immunosorbent assay (ELISA) and then detected for Interleukin-1beta (IL-1beta) in the same sample .Another half (group B) of the sample was used as control for the measurement of Interleukin-1beta (IL-1beta). Another sample of GCF was collected from nine patients with abutment and non-abutment because of the distal extension teeth lost resulted from the chronic periodontitis. At first, one half (group C) of the sample was detected for tumor necrosis factor-alpha (TNF-alpha ) by ELISA and then measured for elastase in the same sample. Another half (group D) of the sample was used as control for the measurement of elastase. RESULTS: There was no significant difference of both the measured value of IL-1beta absorbency (group A and B are 0.5 +/- 0.4 and 0.5 +/- 0.4, respectively, P=0.136)and that of elastase absorbency (group C and D are 1.1 +/- 0.6 and 1.1 +/- 0.6, respectively, P=0.943) between the original elution and the sample after detection of IL-6 or TNF-alpha in the test 1 and test 2, respectively. Moreover, the values showed high correlation (r=0.993, P=0.000 in the group A and B; r=0.979, P=0.000 in the group C and D). CONCLUSION: It is practicable that one sample of GCF can be reused to examine for several components of GCF.