【摘要】 目的 明确γ干扰素释放实验（IGRA）样本处理中的主要影响因素，以减少干扰，促进其临床应用。方法 采集2022年1-12月于广州市胸科医院住院的40例结核患者新鲜抗凝血样本，经外周血与结核刺激物的共培养后，ELISA测试血浆γ干扰素（IFN-γ）水平。采用单因素对照方法，比较样本刺激前保存时间（6、8、10、12、14、16和18 h）、孵育温度（25、30、35、37和39℃）和孵育时间（6、12、18、20、22和24 h），以及孵育后存储（温度15℃）时间（0、24、48和72 h）对IGRA的影响。结果 延长外周血样本的刺激前保存时间时间至18 h对检验结果没有影响；在孵育温度对结果影响研究中发现，低于30℃时对结果的影响差异有统计学意义（P=0.013），在35与37℃的结果差异无统计学意义（P>0.05），在39℃下，IFN-γ浓度下降；在孵育时间对结果影响研究中发现，最佳孵育持续时间为18～22 h；在孵育后样本存储在15℃的时间长度对结果的影响中发现，24、48 h和对照之间的结果差异无统计学意义（P>0.05），而72 h与对照之间的差异有统计学意义（P=0.003）。结论 明确样本在刺激前保存时间可达18 h,37℃孵育20 h和在15℃下可存储48 h，可简化IGRA方法在结核临床诊断中的应用。更多还原

【Abstract】 Objective This study aims to identify the primary influencing factors in the handling of interferon-gamma release assay(IGRA) samples in order to reduce interference and promote their clinical application.Methods Fresh anticoagulated blood samples were collected from 40 hospitalized tuberculosis patients at the Guangzhou Chest Hospital between January and December 2022.After co-culturing with peripheral blood and tuberculosis stimulants,the plasma interferon-gamma(IFN-γ) levels were tested using ELISA.A single-factor control method was used to compare the impact of sample retention time before stimulation(6,8,10,12,14,16 and 18 h),incubation temperature(25,30,35,37 and39℃),incubation time(6,12,18,20,22 and 24 h),and post-incubation storage time(0,24,48 and 72 h) on IGRA.Results Prolonging the retention time before stimulation of peripheral blood samples to 18 h did not affect the test results.When investigating the influence of incubation temperature,a statistically significant difference was found for temperatures below 30℃(P=0.013).There was no statistically significant difference in results between 35 and 37℃(P>0.05),but at 39℃,IFN-γ concentration decreased.In the study on the influence of incubation time,the optimal incubation duration was found to be between 18 and 22 h.Regarding the effect of the length of time the samples were stored at 15℃ after incubation,no statistically significant difference was observed between 24,48 h,and the control(P>0.05),but there was a statistically significant difference between 72 h and the control(P=0.003).Conclusion It has been determined that samples could be stored for up to 18 h before stimulation,incubated for 20 h at 37 ℃,and stored for 48 h at 15 ℃,which simplifies the clinical application of the IGRA method in tuberculosis diagnosis.更多还原