【摘要】 目的评价基于磁珠法的荧光定量PCR检测HBV-DNA的临床应用。方法选用磁珠法定量检测试剂和煮沸法定量检测试剂,对一系列临床患者血清标本进行检测,比较两种试剂检出率的差异;通过浓度为1×108的样本的梯度稀释结果考察两者的灵敏度和线性范围。结果 68例临床样本中,磁珠法试剂检测的阳性率为69.12%,煮沸法试剂检测的阳性率为32.35%(P<0.05);两种试剂对103IU/m L阳性样本检测结果:y=0.913x-0.261,r=0.919;磁珠法线性范围3.9×(101~108),灵敏度39 IU/m L,煮沸法线性范围2.4×(102~107),灵敏度240 IU/m L。结论磁珠法核酸提取试剂线性范围宽,灵敏度高,临床检出率明显高于煮沸法,对于高浓度和低浓度样本都能准确的定值,适合于乙肝治疗后监测与体检筛查。更多还原

【Abstract】 Objective To evaluate magnetic bead-based fluorescent quantitative PCR detection of HBV-DNA in clinical application. Methods Immunomagnetic beads and quantitative detection and boiling method were used to detect a series of patient serum samples. The detection rates of the two detection methods were compared; their sensitivities and linear ranges were determined with the gradient dilutions of samples at the concentration of 1 ×108. Results Of the total 68 clinical samples,the positive rate of bead method was 69. 12%,while that of boiling method was 32. 35%( P < 0. 05). The detection results of the 103 IU / m L positive sample by the two methods were y = 0. 913x- 0. 261,r = 0. 919. The magnetic bead method showed a linear range of 3. 9 ×( 101-108) and a sensitivity of 39 IU / m L,while the boiling method showed a linear range of 2. 4 ×( 102-107) and a sensitivity of 240 IU / m L. Conclusion Magnetic beads method of nucleic acid extraction has wider linear range,higher sensitivity and higher clinical detection rate than the boiling method,and is accurate for both high and low concentration samples,which is suitable for post-treatment monitoring of hepatitis B and physical exam screening.更多还原