荧光定量PCR检测HCVRNA的测量不确定度评定及应用  

Detection of HCV RNA PCR Fluorescence Quantitative Measurement and Application of Uncertainty

作　　者：裴兵[1] 夏寿扬[1] 吴辉[1] 李续亮[2] 

机构地区：[1]江苏省泗洪县人民医院,江苏泗洪223900 [2]山西省肿瘤医院,太原223500

出　　处：《现代检验医学杂志》2011年第3期65-67,71,共4页Journal of Modern Laboratory Medicine

摘　　要：目的对荧光定量PCR测定HCVRNA的测量不确定度进行评定及临床应用。方法利用罗氏LightCycler1．0荧光定量PCR分析仅选取2个HCVRNA结果阳性的临床样本，HCVRNA拷贝数分别为1×10^3 copies／ml～1×10^5 copies／ml和1×10^4 copies／ml～1×10^7 copies／ml，各重复测定10次，对测定HCVRNA潮量过程的分析，确定并简化测量不确定度的来源，采用不确定度A类评定方法以及不确定度B美评定方法，量化各不确定度分量，确定合成不确定度与扩展不确定度。结果测量不确定度来源主要有：批内重复性、批间重复性，温度时间，消毒前消毒后和方法偏倚等因素。在各分量中，由温度时间影响导致的不确定度所占比例最大，批间重复性试验所占比例最小。荧光定量PCR测定HCVRNA的扩展不确定度U=0．268。结论通过荧光定量PCR测定HCVRNA测量不确定的研究有助于各实验室结果的表达趋于一致，可使来自不同实验室的结果或同一实验室不同时期的结果具有可比性。Objective Determination of HCV RNA PCR fluorescence quantitative measurement uncertainty evaluation and clinical application. Methods Fluorescence quantitative PCR using Roche LightCyclerl. 0 analyzer results selected two HCV RNA positive clinical samples,HCV RNA copy number was 1 × 10^3 -1× 10^5 copies/ml and 1 × 10^5 -1× 10^7 copies/ ml. Repeated measurements 10 times,on the determination of HCV RNA analysis of the measurement process,and simplified the measurement uncertainty to determine the source,the use of class A uncertainty evaluation method and type B uncertainty evaluation method to quantify the uncertainty components ,determined the synthesis uncertainty and expanded uncertainty. Results The main sources of uncertainties of measurement were：within run variance ,run to run variance ,temperature and time influence,contrast disinfect with no disinfect and method bias. The uncertainty caused by was temperature and time found mostly and precision between laboratory least. The expanded uncertainty of HCV by FQ-PCR was 0. 268. Conclusion Expanded uncertainty could be compared in different results of HCV by FQ-PCR and it provides guide significance observing the cure effect of anti-HCV and choosing the concentration of quality control.

关 键 词：荧光定量PCR 丙肝病毒核酸 测量不确定度 

分 类 号：Q503[生物学—生物化学]