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目的分析时间分辨荧光免疫分析法(TrFIA)检测干血滤纸片17α-羟孕酮(17α-OHP)的影响因素并提出对策,以提高新生儿先天性肾上腺皮质增生症(CAH)的筛查质量。方法将干血滤纸片标本分别按抗血清和铕示踪剂的加样量(150μL、110μL、100μL、90μL、50μL)分5组,采用TrFIA法测定干血滤纸片17α-OHP的浓度;比较58孔微孔板底部去除水汽前、后的17α-OHP浓度;比较30例干血斑不同取样部位(中央和边缘)的17α-OHP浓度。结果抗血清和铕示踪剂的加样量对测定结果的影响与100μL组比较,差异有统计学意义(P<0.01);微孔板底部水汽存在与否对检测结果的影响有统计学意义(P<0.01);取样部位(中央和边缘)对测定结果影响不大,差异无统计学意义(P>0.05)。结论新生儿CAH筛查试验时应保证试剂加样量准确,微孔板底部干燥。
Objective To analyze the influencing factors of 17α-hydroxyprogesterone (17α-OHP) in dry blood filter paper by time-resolved fluorescence immunoassay (TrFIA) and to propose countermeasures to improve the quality of screening of congenital adrenal hyperplasia (CAH) in neonates . Methods The dry blood filter paper samples were divided into 5 groups according to the amount of antiserum and europium tracer (150μL, 110μL, 100μL, 90μL, 50μL) respectively. The concentration of 17α-OHP was determined by TrFIA method. 17α-OHP concentration before and after the water vapor was removed from the bottom of the 58-well microplate; and 17α-OHP concentrations were compared among 30 samples (central and peripheral) of the dried blood spots. Results The effect of antisera and europium tracers on the determination results was statistically significant compared with 100μL group (P <0.01). The presence or absence of water vapor at the bottom of microplate had a significant effect on the detection results (P <0.01). There was no significant difference between the sampling sites (center and periphery) and the difference between the two groups (P> 0.05). Conclusion Neonatal CAH screening test should ensure that the reagent loading volume accurately, the bottom of the microplate dry.