【摘要】 目的探讨聚合酶链反应(PCR)检测全血中乙型肝炎病毒(HBV)DNA含量及临床应用价值。方法取20μl血清裂解液提取HBVDNA、和取同样量的全血经不同裂解液提取血液中血清及白细胞内总HBVDNA,同时经PCR检测。对78份乙型肝炎表面抗原(HBsAg)阳性患者两种取材方法检测HBVDNA含量进行比较。结果检测HBVDNA的最低检测限为1×103拷贝/ml,29份乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBeAg)和抗乙型肝炎核心抗体(抗HBc)均为阳性患者全血检测HBVDNA阳性率为100%,血清阳性率为100%,P>0.05;37份HBsAg、抗乙型肝炎e抗体(抗HBe)和抗HBc均为阳性患者全血检测HBVDNA阳性率为97.30%,血清阳性率为48.65%,P<0.01;12份HBsAg和抗HBc均为阳性患者全血检测HBVDNA阳性率为91.67%,血清阳性率为25%,P<0.01;20份非乙型肝炎患者血清均为阴性。血清HBVDNA阳性多数是ALT升高患者,其拷贝数均小于全血测定的拷贝数。结论全血检测HBVDNA阳性率高,避免血液中白细胞内HBV漏检,它能准确地反应血液HBV含量,更好地为临床治疗效果作监测。更多还原

【Abstract】 Objective To evaluate the clinical application value of determination of the content of HBV DNA in human whole blood. Methods HBV DNA from 20μl serum which was lysed in cell lysis buffer and from 20μl human whole blood which was lysed in several different cell lysis buffers was extracted. Both of them were detected with PCR.Comparing the HBV DNA content extracted with the two methods of 78 cases with HBsAg positive. Results The lowest limit for detection of HBVDNA was 1×103 copies. Positive rate of HBV DNA dectected from the human whole blood group was 100% of 29 cases with HBsAg,HBeAg and anti-HBc positive, and that of serum group was 100%(P>0.05). Positive rate of HBV DNA detected from human whole blood group was 97.30% of 37 cases with HBsAg,anti-HBe and anti-HBc positive,but that of serum group was 48.65%(P<0.01). Positive rate of HBV NDA dectected from the human whole blood group was 91.67% of 12 cases with HBsAg and anti-HBc positive,but that of serum group was 25%. The rate of 20 normal serum were negative(P<0.01). ALT were increased in most HBV DNA positive serum, all its copies were smaller than the detective copy number in whole blood. Conclusions The positive rate of HBV NDA in human whole blood group is higher than that in serum group. To detect HBV DNA from whole blood can avoid missing any HBV DNA within leukocytes. It is suitable for detecting HBV DNA content and helpful for surveillance of clinical therapeutic effects.更多还原