两种核酸提取法对不同程度溶血标本HBV-DNA检测结果的比较

目的探讨一步核酸提取法和两步核酸提取法对不同程度溶血标本乙型肝炎病毒(HBV)DNA检测结果影响的差异。方法收集22例临床随机患者全血标本,离心并于当天用一步和二步核酸提取法试剂检测HBV-DNA含量后,将全血标本放4℃冰箱保存,每隔2~3 d取出分别用两种试剂检测HBV-DNA的含量。结果 22例标本随着存放时间和溶血程度的增加,其HBV-DNA含量有下降趋势;一步法在标本存放3 d后结果显著低于首次检测结果(t3d=3.981,P<0.01);二步法在标本存放19 d后结果显著低于首次检测结果(t3d=3.642,P<0.01);标本于4℃存放3天后一步法HBV-DNA的检测结果显著低于二步法(t3d=4.91,P<0.001)。结论溶血标本对二步核酸提取法试剂检测HBV-DNA含量的影响作用远低于一步法,对于临床由于特殊原因无法及时获得合格血标本时,可考虑使用二步核酸提取法试剂检测HBV-DNA的含量。

Research on the detection of HBV-DNA in different hemolysis sample by two kinds of nucleic acid extraction methods

Objective To discuss the difference between one-step nucleic acid extraction and two-step nucleic acid extraction for HBV-DNA in different degrees of hemolysis sample.Methods 22 Whole blood samples were collected from clinical patients randomized,centrifuged and we detected the content of HBV-DNA by one-step and two-step PCR method seperately on that day.With 4℃ preservation,the content of HBV-DNA was testd every 2-3 days using two reagents.Results As the storage time and hemolysis increased,the contents of HBV-DNA decreased.The HBV-DNA content of samples preserved 3 days detected by one-step significantly was lower than that of the first detection（td=3.981,P0.01）,in the same way,the HBV-DNA content of samples preserved in 19 days detected by two-step significantly was lower than that of the first detection（td=3.642,P0.01）.Morever,the HBV-DNA content of samples preserved 3 days in 4 ℃ detected by one-step was significantly lower than that of two-step（t=4.91,P0.001）.Conclusion The effect of hemolysis sample on the detection of HBV-DNA by two-step method is more lower than that of one-step method,It is better to use two-step method to detect HBV-DNA in samples which is not acquired normally in some cases.