中文摘要：

目的探讨PCR仪温度特性及变性温度、变性时间对实时荧光定量PCR检测乙型肝炎病毒的影响。方法分析比较变性温度范围为88℃～97℃,变性时间分别为5s和40s,退火和延伸60℃,40s,循环数40的实时荧光定量PCR检测乙型肝炎病毒的效果。结果变性温度超过97℃或者变性时间超过40s,检测到的DNA的量明显降低;变性时间小于20s,模板含量较低的样本难以检出。结论变性初期或退火初期PCR温度与程序设置温度偏差较大;变性时间不应超过95℃,否则,检测到的DNA数量将显著降低;变性时间不应小于20s或超过40s,否则容易导致假阴性。

英文摘要：

【Objective】Effect of thermal characteristics on PCR processors, denaturation temperature, denaturation time on the validity of detecting HBV-DNA （150 bp amplified） were studied by real-time fluorescent quantification PCR. 【Method】Denaturation are from 88 ℃ to 97 ℃ for 5 seconds and 40 seconds respectively, annealing at 60 ℃ for 40 seconds; 40 cycles. 【Results】When denaturation tempertaur exceeds 97 ℃ or denaturation time is longer than 40 seconds DNA numbers tested decrease remarkably. False negative would be induced if denaturation time is longer than 40 seconds or lower than 20 seconds.