环境温度对Luminex流式荧光免疫分析法测定糖类抗原242结果的影响

Effect of ambient temperature on determination of carbohydrate antigen 242 by Luminex flow cytometry fluorescence immunoassay

目的分析定标和测定时不同环境温度下Luminex流式荧光免疫分析法测定糖类抗原242(CA242)结果的差异,评价环境温度对测定结果的影响。方法选择高值和中值2个不同浓度的标本,分别采用Luminex流式荧光免疫分析法在11个温度点(18.3、20.1、21.5、22.2、24.1、25.0、25.4、26.5、27.4、28.5、30.0℃)重复测定CA242 5次,以定标时环境温度(25℃)下的检测结果的平均值为参比值,计算不同环境温度下检测结果的偏倚。随机选择49例涵盖高、中、低值的标本,分别采用Luminex流式荧光免疫分析法在20、25和30℃下对CA242进行测定,比较测定结果并分析其相关性。结果中值标本和高值标本在18.3℃下测定结果的偏倚分别为-35.6%和-29.4%。49例标本在20℃下的测定结果与25℃下的测定结果差异有统计学意义(P<0.001),并且二者具有相关性(回归方程:Y=0.676 9X+0.374 7,R2=0.990 5);在30℃下的测定结果与25℃下的测定结果差异有统计学意义(P<0.001),并且二者具有相关性(回归方程:Y=0.896 6X+0.227 0,R2=0.999 4)。结论环境温度过低或过高会对Luminex流式荧光免疫分析法测定CA242产生负性影响,测定时应使环境温度保持相对恒定且控制在定标温度±4℃范围内。

Objective To analyze the difference of the carbohydrate antigen 242(CA242) results measured by Luminex flow cytometry fluorescence immunoassay with different calibrations and different ambient temperatures, so as to evaluate the effect of ambient temperature on the measurement results. Methods Two CA242 samples with different concentrations(median-and high-value) were selected and the determination was repeated at 11 temperature points(18.3,20.1, 21.5, 22.2, 24.1, 25.0, 25.4, 26.5, 27.4, 28.5, and 30.0 ℃) for 5 times by Luminex flow cytometry fluorescence immunoassay. The average value of the detection result at the calibration temperature(25 ℃) was taken as the reference value, and the biases of the detection results at different ambient temperatures were calculated. Forty-nine CA242 specimens covering high-, median-and low-values were examined at 20, 25 and 30 ℃ by Luminex flow cytometry fluorescence immunoassay, and the results were compared and correlation analysis was carried out. Results The biases of the medianand high-value samples at 18.3 ℃ were-35.6% and-29.4%, respectively. The difference between the measurement results at 20 ℃ and 25 ℃ was significant for the 49 specimens(P<0.001), and there was a linear correlation between them(Y=0.676 9 X+0.374 7, R2=0.990 5). The difference of measurement results was significant between 30 ℃ and 25 ℃(P<0.001), and there was a linear correlation between them(Y=0.896 6 X+0.227 0, R2=0.999 4). Conclusion Too low or too high ambient temperature has a negative effect on the determination of CA242 by Luminex flow cytometry fluorescence immunoassay. The ambient temperature should be kept relatively constant and controlled within the range of calibration temperature ±4 ℃.