样本处理后静置时间对HBV—DNA荧光定量PCR的影响

目的：探讨乙型肝炎病毒核酸（HBV-DNA）荧光定量聚合酶链反应（FQ-PCR）在样本处理后立即加样扩增和4℃冰箱静置24h后再加样扩增对检测结果的影响。方法：68例不同模式乙肝患者样本分别用2种方法处理后进行HBV-DNAFQ-PCR检测。结果：68例样本采用第1种方法共检出HBV-DNA阳性35例，第2种方法共检出39例；第2种方法HBV-DNA阳性定量结果普遍高于第1种方法，指数平均高1个次方左右（u=5.14，P〈0.05）；以第2种方法检测结果为依据，则第1种方法有4例阳性结果漏检，检测结果假阴性率达10.3％（4／39）。结论：样本处理后，应置4℃冰箱静置一定时间，以保证病毒颗粒充分裂解后再进行HBV-DNAFQ-PCR，以提高HBV-DNA的检出率。

Effects of Standing Time for Disposed Samples on HBV-DNA Fluorescent Quantita tive PCR

Objective： To investigate the effects of immediate amplication of sample as well as amplication for disposed samples after 24 h standing in 4℃ fridge on HBV-DNA detection results by using FQ-PCR method. Methods： To execute HBV-DNA detection on 68 cases of different modes of HB patients by using FQ-PCR method in two different ways. Results： 35 cases with positive HBV-DNA among total 68 cases of samples were detected in the first way, 39 cases with positive HBV-DNA were detected in second way. The quantitative result of positive HBV-DNA detected in second way is universally higher than that in the first way, the average index being on square（u=5.14, P〈0.05）. Based on the second detection result, 4 cases with positive HBV-DNA were left out when the first way was adopted, with false negative rate of HBV-DNA being 10.3%（4/39）. Conclusion： After samples of HBV-DNA FQ-PCR have been disposed, they should be put in 4℃ for a certain time of standing to ensure that virus particles fission completely before they were amplified in order to increase the sample detection rate.