Lin H J, Wu P C, Lai C L, Chak W
Clin Chem. 1984 Apr;30(4):549-52.
A micromethod for the specific measurement of hepatitis B viral DNA polymerase in serum is presented, based on the phosphonoformate inhibition assay (J Med Virol 12: 61-70, 1983). In the micromethod, sample volume is reduced to 120 microL and the ultracentrifugation step is eliminated. The method allows good discrimination between serum infected with hepatitis B virus and uninfected serum. The cutoff value for rate of nucleotide incorporation, based on assays of 41 serum specimens negative for hepatitis B serological markers, was about 15 nU/L (90th percentile). Serum containing hepatitis B surface and antigens exhibited rates of phosphonoformate-inhibitive nucleotide incorporation of 150 (SD 150) nU/L, with an upper 90th percentile range of 17 to 667 nU/L (n = 41). The micromethod makes use of commercially available [32P]dCTP (specific activity about 7000 kCi/mol). 125I-labeled dCTP was found to be unsuitable for this assay. Human DNA polymerases in serum are detected by this method but are excluded from the phosphonoformate-inhibitive fraction.
本文介绍了一种基于膦甲酸抑制试验（《医学病毒学杂志》12: 61 - 70, 1983）的血清中乙型肝炎病毒DNA聚合酶特异性测定的微量方法。在该微量方法中，样本体积减少至120微升，省去了超速离心步骤。该方法能很好地区分感染乙型肝炎病毒的血清和未感染的血清。基于对41份乙型肝炎血清学标志物阴性的血清标本的检测，核苷酸掺入率的临界值约为15 nU/L（第90百分位数）。含有乙型肝炎表面抗原和e抗原的血清，膦甲酸抑制的核苷酸掺入率为150（标准差150）nU/L，第90百分位数上限范围为17至667 nU/L（n = 41）。该微量方法使用市售的[32P]dCTP（比活约7000 kCi/mol）。发现125I标记的dCTP不适用于此检测。该方法可检测血清中的人DNA聚合酶，但它们不包含在膦甲酸抑制部分中。
