Line-Gene实时荧光定量PCR不同结果分析模式对检测结果的影响

Effects of Different Analysis Mode of Line-Gene Real Time Quantitative PCR System on Results

目的探讨实时荧光定量PCR不同结果分析模式对检测结果的影响.方法用Line-Gene FQD-33A型荧光定量PCR检测系统检测本院门诊和病房乙肝病人血清HBV DNA 77份,分别用最大二阶导数法和样点拟合法进行结果分析.结果发现两种分析方法的总体相关性较好(r= 0.978). 但HBV DNA拷贝数含量＜105时, 两种模式的相关性差(r= 0.706),此时,最大二阶导数法得出的数值要比样点拟合法低(t=2.61,P＜0.05),且易出现假阴性结果.结论虽然样点拟合法步骤较多,但其结果更可靠,因此进行结果分析时,建议使用样点拟合法.而且当样本中HBV DNA拷贝数＜105时只能使用样点拟合法进行结果分析.

Objective To observe the effect of different quantitative analysis modes on results. Methods Using line-Gene fluoro-genic quantitative PCR detection system to detect HBV DNA of 77 patients and second derivative maximnm method and fit point method to analyze results. Results In general, results got from the two methods had good relativity( r = 0.9789 ). But when HBV DNA copy is less than 105 per ml, the relativity is poor( r = 0.706 ). Results got from second derivative maximum method is lower than from fit point method, and prone to get false negative results. Conclusion Although fit point method is much complicated than second derivative maximum method, the corresponding results is more reliable. Fit point method should be the first choice, and when DNA copy is less than 105 per ml, fit point method is the only method one could use.