中性粒细胞化学发光及血清调理功能测定的方法学研究

Study on the measurement method of PMN CL and serum opsonic capacity

目的建立中性粒细胞化学发光及血清调理功能测定的最佳实验体系。方法通过探讨分离的人外周血中性粒细胞化学发光（ＰＭＮ－ＣＬ）测定的最佳实验条件及若干影响因素，建立最佳实验体系。血清调理功能测定酵母多糖系用自体血清调理，ＰＭＮ－ＣＬ开始至出现高峰的时间作为血清调理功能的活性分析。结果ＰＭＮ－ＣＬ测定的最佳反应体系为：反应温度３７℃±０．５℃，ＰＭＮ悬液（１×１０９Ｌ－１）１ｍｌ，鲁米诺溶液（２．５×１０－５ｍｏｌＬ－１）０．２ｍｌ，调理的酵母多糖悬液（１０ｇＬ－１）０．２ｍｌ，分离的ＰＭＮ悬液置４℃保存５ｈ内，ＣＬ强度基本不变。

Objective To set up the optimal experimental system for chemiluminescence of neutrophils and for serum opsonic capacity. Methods In order to set up the optimal experimental system, We have examined the experimental conditions and some influencing factors to chemiluminescence of human peripheral neutrophils (PMN CL). Autosera was used to measure zymosan in serum opsonic capacity test system. The time point of peak volume of CL was applied for analysis of serum opsonic capacity. Results The optimal experimental conditions for measurement of PMN CL are reaction temperature at 37℃±0 5℃. Reaction system is a mixture of PMN suspension 1 ml at concentration 1×10 9·L -1 , luminol solution 0 2 ml at concentration 2 5×10 -5 mol·L -1 and opsonized zymozan suspension 0 2 ml at concentration 10 g·L -1 . The CL of freshly isolated PMN suspension is stable within 5 hours at 4℃. Conclusion Conditions of PMN CL and serum opsonic capacity may be the indication of human immunity.