干滤纸血片制备环境的理化因素对实验结果的影响

Effects of physical and chemical factors in environment for dried blood spot preparation of neonatal screening assay

目的观察新生儿疾病筛查干滤纸血片(dried blood spots)制备环境中显著影响实验结果的理化因素。方法研究组1:收集正常标本60例、阳性标本120例,于对照条件及10种特定环境下制备干滤纸血片。研究组2:另收集正常标本30例、阳性标本80例,于对照条件及7种甲醛浓度环境下制备干滤纸血片。用荧光法或时间分辨荧光免疫分析法检测标本中苯丙氨酸(Phe)、葡萄糖-6-磷酸脱氢酶(G6PD)、促甲状腺激素(TSH)、17α-羟孕酮(17α-OHP)水平。用SPSS 22.0软件进行统计学分析。结果 10种环境下制备的干滤纸血片Phe检测结果与对照组差异无统计学意义(P>0.05),甲醛敏感阈为4.62~6.95 ppm/18 h;37℃烘箱、潮湿、紫外线照射、新装修环境、乙醇、冰醋酸、甲醛等环境下制备的干滤纸血片G6PD检测结果低于对照组(P<0.05),甲醛敏感阈为0.30~0.38 ppm/4 h及0.21~0.24 ppm/18 h;潮湿、紫外线照射、甲醛环境下制备的干滤纸血片TSH检测结果低于对照组(P<0.05),甲醛敏感阈为0.32~0.52 ppm/4 h及0.38~0.45 ppm/18 h;潮湿、紫外线照射、甲醛环境下制备的干滤纸血片17α-OHP检测结果低于对照组(P<0.05),甲醛敏感阈为4.37~4.62 ppm/4 h及0.38~0.45 ppm/18 h。冷风快速吹干与2~8℃冷库环境下制备干滤纸血片4项检测结果与对照组差异均无统计学意义(P均>0.05)。结论干滤纸血片制备环境的理化因素与新生儿疾病筛查的准确性密切相关,甲醛、乙醇、冰醋酸、紫外线照射、高温、潮湿及装修污染等是干滤纸血片制备环境的必要管控因素;冷风快速吹干与2~8℃冷库过夜可作为新生儿筛查干滤纸血片制备的备选方法。

Objective To investigate the effects of physical and chemical factors in the environment for dried blood sample （DBS） preparation of neonatal screening assay. Methods A total of 60 normal and 120 positive DBS were prepared under control and 10 different conditions. Another 30 normal and 80 positive DBS were prepared under control and 7 different concentration gradients of formaldehyde. The levels of phenylalanine （Phe）, glucose-6-phosphate dehydrogenease （G6PD）, thyroid stimulating hormone （TSH） and 17α-hydoxyprogesterone （17α-OHP） were tested by time-resolved fluorescence immunoassay or fluorescence assay. Statistical analysis was performed using SPSS 22.0 software. Results Compared with the control group, the results of Phe were not significantly different （P 〉0.05） when the samples were dried under the formaldehyde sensitive threshold （4.62 to 6.95 ppm for 18 hours）. G6PD levels were significantly lowered when the samples were dried under all the conditions except for fast cold drying （2 to 8 ℃ overnight and formaldehyde condition, 0.30 to 0.38 ppm for 4 hours or 0.21 to 0.24 ppm for 18 hours）. TSH and 17α-OHP levels were lowered obviously when the samples were dried under the conditions of humidity, UV and formaldehyde condition （ TSH ： 0.32 to 0 52 ppm for 4 hours, 0.38 to 0.45 ppm for 18 hours, 17α-OHP： 4.37 to 4.62 ppm for 4 hours, 0.38 to 0.45 ppm for 18 hours）. The results of Phe, G6PD, TSH and 17α-OHP were not statistically different with the control group when the samples were dried under the fast cold drying and 2 to 8 ℃ overnight. Conclusion The physical and chemical factors in the environment of DBS preparation should be relat- ed to the accuracy of neonatal disease screening closely. The necessary control factors including formaldehyde, ethanol, glacial acetic acid, ultraviolet irradiation, heat, humidity and decoration pollution may exhibit significant effects on the preparation of DBS. Fast cold drying and overnight at 2 to 8 ℃ could be available for DBS preparation.