不同的样本处理方法对乙型肝炎病毒DNA检测的影响

Effect of varied sample treatment procedures on detection of hepatitis B virus DNA

目的探讨乙型肝炎病毒（ＨＢＶ）的聚合酶链反应（ＰＣＲ）常规检测中血清样本的最佳处理方法。方法分别用不同缓冲液（ＰＢＳ与ＴＥ）对血清进行１：１或１：１０稀释，不同浓度去污剂（ＴｒｉｔｏｎＸ－１００与ＮＰ４０）及巯基乙醇处理血清样本，以及用硫氰酸胍－酚－氯仿抽提、蛋白酶Ｋ消化、碱裂解、辛酸钠裂解等方法处理血清样本，比较这些方法对ＰＣＲ法检测血清ＨＢＶＤＮＡ敏感度的影响，并进行了辛酸钠最适浓度实验。结果在所比较的１８种方法中发现硫氰酸胍－酚－氯抽提法、碱裂解法及辛酸钠法有较高的检测敏感度，而辛酸钠法有更好的重复性，方法最为简便。

The detective sensitivity vary obviously in the assay of HBV DNA by means of polymerase chain reaction with 18 different sample treatment procedures.Higher sensitivity was found in three methods, namely,guanidine thiocyanate/phenol/chloroform,NaoH denaturation and sodium octanoate.The procedure of phenol/chloroform was too labor-intensive to be accepted in routine clinical testing, sodium octanoate which may suppress the inhibitory effect of denatured albumin on the polymerase chain reaction and thus the sensitivity was higherthan the method of NaOH denaturation.Treatment of samples with 30～50mmol/L of sodium octanoate in final concentration for the detection of HBV DNA has shown to be very sensitive, simple and reproducible.