猪全血法体外检测热原的可行性研究

目的：研究猪全血法体外检测热原的可行性。方法：采用猪全血，与不同浓度的各类热原 共同孵育20 h（37 ℃、5%CO₂），以细胞因子（IL-1β、IL-6）为检测指标，用酶联免疫法（ELISA）检测孵育体系中相应细胞因子的含量，进行猪全血-IL-1β、IL-6法体外热原检测的方法研究。结果：猪全血经一定比例稀释后（猪全血：RPMI1640为1：3），猪全血-IL-1β、IL-6法对LPS可检测到0.25 EU·mL^-1，对LTA可检测到0.5 μg·mL^-1，对酵母多糖可检测到16.0 μg·mL^-1；且在该孵育体系下，猪全血-IL-1β法对LPS的最低检测限为0.125 EU·mL^-1，线性范围为0.125~0.5 EU·mL^-1，r=0.9515；猪全血-IL-6法对LPS的最低检测为0.0625 EU·mL^-1，线性范围为0.0625~0.5 EU·mL^-1，r=0.9926。结论：初步研究认为，猪全血-IL-1β、IL-6法适用于检测热原。

Objective: To explore feasibility of porcine whole blood for in vitro detection of pyrogen. Methods: The porcine whole blood mixed with different levels of pyrogen was incubated for 20 h (37 ℃, 5% CO₂). With cell factors (IL-1 β, IL-6) as indicators of detection, enzyme-linked immunosorbent assay (ELISA) was employed for the content of corresponding cytokine in the incubation system, and the feasibility of porcine whole blood with IL-1 β,IL-6 method for in vitro pyrogen detection was explored. Results: Porcine whole blood diluted by certain proportion (porcine whole blood:RPMI1640 to 1:3), porcine whole blood-IL-1β, IL-6 method could detect 0.25 EU·mL^-1 of LPS, 0.5 μg·mL^-1 of LTA, 16 μg·mL^-1 of Zymosan; and in the incubation system, the detection limit of LPS by the method with porcine whole blood-IL-1β was 0.125 EU·mL^-1;the linear range was 0.125-0.5 EU·mL^-1, r=0.9754; the minimum detection with porcine whole blood-IL-6 method for LPS was 0.0625 EU·mL^-1;the linear range was 0.0625-0.5 EU·mL^-1, r=0.9959. Conclusion: The preliminary study concluded that porcine whole blood-IL-1β, IL-6 method is suitable for the detection of pyrogen.