Knöll Antje, Louwen Frank, Kochanowski Bernd, Plentz Annelie, Stüssel Julia, Beckenlehner Karin, Jilg Wolfgang, Modrow Susanne
Institute of Medical Microbiology and Hygiene, University of Regensburg, Germany.
J Med Virol. 2002 Jun;67(2):259-66. doi: 10.1002/jmv.2216.
Human parvovirus B19 infections are common in the general population, and infection during pregnancy may cause hydrops fetalis and fetal death. To initiate adequate treatment, accurate laboratory diagnosis is essential. The most sensitive tests are nested PCR systems, but these assays provide semiquantitative results at best. A parvovirus B19 DNA assay was developed based on the real time TaqMan PCR. This method was calibrated on the basis of serial plasmid dilutions and tested with an international parvovirus B19 standard. The assay was capable of quantifying parvovirus B19 DNA from one to about 5 x 10(7) genome equivalents per reaction (corresponding to 100 to 5 x 10(9) genome equivalents per ml serum). Samples from 51 pregnant women with suspected acute parvovirus B19 infection were tested, and positive PCR results were obtained in at least one of the materials investigated in 41 cases. The median viral DNA load in maternal blood samples was 1.3 x 10(4) copies/ml (range 7.2 x 10(2)-2.6 x 10(7)). Maternal virus DNA concentration was not associated with the presence of maternal symptoms and/or fetal complications. As the stage of infection was not known in the majority of cases, our data do not exclude an association between peak levels of parvovirus B19 DNA and the development of complications. Maternal sera and corresponding fetal material were available for concurrent testing from 15 DNA-positive cases: in most fetal samples, viral DNA concentrations were several orders of magnitude higher (up to 2.1 x 10(12) copies/ml) compared to the corresponding maternal blood samples.
人细小病毒B19感染在普通人群中很常见，孕期感染可能导致胎儿水肿和胎儿死亡。为了开展适当的治疗，准确的实验室诊断至关重要。最敏感的检测方法是巢式PCR系统，但这些检测最多只能提供半定量结果。基于实时TaqMan PCR开发了一种细小病毒B19 DNA检测方法。该方法基于系列质粒稀释进行校准，并用国际细小病毒B19标准品进行检测。该检测方法能够对每个反应中1至约5×10⁷个基因组当量的细小病毒B19 DNA进行定量（相当于每毫升血清中100至5×10⁹个基因组当量）。对51例疑似急性细小病毒B19感染的孕妇样本进行检测，在41例所调查的样本中至少有一份获得了阳性PCR结果。孕妇血液样本中病毒DNA载量的中位数为1.3×10⁴拷贝/毫升（范围为7.2×10²至2.6×10⁷）。孕妇病毒DNA浓度与孕妇症状和/或胎儿并发症的存在无关。由于大多数病例的感染阶段未知，我们的数据并不排除细小病毒B19 DNA峰值水平与并发症发生之间的关联。从15例DNA阳性病例中可获得孕妇血清和相应的胎儿样本进行同步检测：在大多数胎儿样本中，病毒DNA浓度比相应的孕妇血液样本高出几个数量级（高达2.1×10¹²拷贝/毫升）。
