摘要
目的:建立核酸检测技术(NAT)用于血液筛查的室内质量控制(QC)方法。方法:将浓度为60 IU/ml HBV-DNA质控标准品,以6个混样常规筛查浓度稀释至10.0 IU/ml,接近检测下限的2倍。如出现混样结果阳性,则需进行拆分单检,以同管HBV-DNA质控品(浓度为60 IU/ml)作为拆分单检质控品,与需拆分样品同时单检。分别通过平行检测各20份同一批次质控品的方式,所得结果用即刻法分析其核酸扩增循环数,计算Ct值的均值()、标准差(SD)和变异常数(CV),以±2SD为警告限,超过±3SD为失控限,输入Excel后绘制LeveyJennings质控图,并以此分别建立核酸筛查混样和拆分单检室内质控图框架。结果:混样和拆分单检连续20次检测结果,其±2SD分别为33.03±1.47和30.08±0.98;±3SD分别为33.03±2.20和30.08±1.47;CV分别为2.22﹪和1.63﹪。t检验计算其P值分别为0.08和0.17。在此质控标准下,在同等条件下混样检测60份、拆分单检13份质控品,检测结果为混样检测59次结果在控,1次失控;拆分单检13次全部在控。阴阳性对照物结果均符合实验要求,结果有效。结论:核酸筛查时混样检测和拆分单检采用不同浓度的质控品,混样时以接近试剂检测限的弱阳性血清作为核酸筛查的质控品,拆分单检时的浓度是混样检测浓度的6倍。此质控方法可保证检测核酸提取和扩增的有效性,提高实验结果的稳定性。
Objective: To establish and evaluate the methods of internal quality control in blood donor screening by nucleic acid test( NAT). Methods: After HBV- DNA standard quality control( QC) sample( 60 IU / ml) w as diluted by pooling 6 samples,the concentration w as 10. 0 IU / ml,w hich w as approach tw ice of the low limit. When the pooling result turned out reactive,the pooling samples need to be split into single sample to process. M eanw hile,the standard QC samples w ere tested as w ell. The same batch QC samples w ere tested 20 times respectively,calculate the mean( ),standard deviation( SD) and CV. M ake Levey- Jennings QC curves by setting ± 2SD as w arning,± 3SD as rejected.The Levey- Jennings quality controls chart w as mapped by using M icrosoft Excel. Results: After 20 times test of mixed / split samples,the ± 2SD w ere 33. 03 ± 1. 47 and 30. 08 ± 0. 98,the ± 3SD w ere 33. 03 ± 2. 20 and 30. 08 ±1. 47,the CV w ere 2. 22% and 1. 63%,respectively. The P value of t test w as 0. 08 and 0. 17 respectively,there w as no statistically significant difference betw een the 2 group. Conclusion: When establish an internal QC system in the screening laboratory by nucleic acid testing,the concentration of the QC samples should be equal to normal specimens.This type of QC system may validate the extraction and amplification of the nucleic acid,and improve the stability of the test results.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2016年第3期921-924,共4页
Journal of Experimental Hematology
