自制大容量扣式负压抽提装置应用于RNA抽提的性能评价

目的对自制大容量扣式负压抽提装置进行扩增前性能评价。方法采用配套负压抽提装置(对照组装置)和自制大容量扣式负压抽提装置(实验组装置)对67份白细胞(WBC)大于2.0×109/L的新鲜全血进行核酸(RNA)提取并进行扩增前性能评价,验证实验组的工作效率、提取RNA的浓度、纯度及完整性。结果 67份RNA标本采用对照组装置69min完成抽提,处理效率平均为0.97份/分;实验组装置41min完成抽提,效率为1.63份/分。对照组抽提标本RNA浓度为(248.8±21.4)μg/mL,实验组为(260.3±21.8)μg/mL。对照组抽提标本OD260/OD280和OD260/OD230分别为(1.995±0.095)和(2.020±0.082),95%CI分别为1.964~2.025和2.001~2.040,实验组分别为(2.093±0.092)和(2.071±0.120),95%CI分别为2.075~2.113和2.044~2.103;2组比较差异有统计学意义(t=24.570,P0.001),线性回归方程Y=0.950 X+0.039,R2=0.903。2组装置提取RNA后完整性均较好,电泳条带整齐、清晰。结论自制大容量扣式负压抽提装置具有更高的抽提工作效率,且能保证RNA浓度、纯度及完整性,更符合实验室实际情况,具有更优的临床价值。

Performance evaluation of the one self-developed device for transmitting negative pressure in RNA extraction

Objective To evaluate the self‐developed device for transmitting negative pressure with large capacity before propa‐gating .Methods Totally 67 ribonucleic acid(RNA) of WBC > 2 .0 × 109 /L fresh whole blood were extracted with the intrinsic de‐vice(control) and the self‐developed device for transmitting negative pressure with large capacity(test) in parallel .The difference of the performance including efficiency ,concentration ,purity and integrity of RNA extraction were evaluated between the two groups . Results In 67 specimens of RNA extraction ,efficiency ,concentration ,purity and integrity of the control device were 0 .97(portion/minute) ,(248 .8 ± 21 .4)μg/mL ,(1 .995 ± 0 .095) (OD260/OD280) ,(2 .020 ± 0 .082) (OD260/OD230) ,1 .964 - 2 .025 (95% con‐fidence interval for mean) (OD260/OD280) ,2 .001 - 2 .040 (95% confidence interval for mean) (OD260/OD230) and complete . Those of test device were 1 .63 portion/minutes ,(260 .3 ± 21 .8)μg/mL ,(2 .093 ± 0 .092) at OD260/OD280 ,(2 .071 ± 0 .120) at OD260/OD230 ,2 .075 - 2 .113 for 95% confidence interval for mean at OD260/OD280 ,2 .044 - 2 .103 for 95% confidence interval for mean at OD260/OD230 and complete .The t value was 24 .570 (P< 0 .001) in paired t‐test ,and the linear regression equation was Y = 0 .950X + 0 .039 ,R2 = 0 .903 for the two groups data of RNA concentration .Conclusion The self‐developed device for transmitting negative pressure with large capacity is excellent in the work efficiency ,concentration ,purity ,integrality and of the RNA in extracting .It is better than the intrinsic device in the clinical value for situation of lab .