Patou G, Pillay D, Myint S, Pattison J
Department of Medical Microbiology, University College & Middlesex School of Medicine, London, United Kingdom.
J Clin Microbiol. 1993 Mar;31(3):540-6. doi: 10.1128/jcm.31.3.540-546.1993.
The characterization and application of a nested polymerase chain reaction (PCR) assay for the detection of human parvovirus B19 DNA is described. The assay was evaluated with 149 diagnostic serum samples (collected up to 150 days after the onset of symptoms) previously tested by dot blot hybridization for B19 DNA and by class-specific capture radioimmunoassays for the detection of B19 immunoglobulin M (IgM) and IgG. B19 DNA was detectable by the PCR in 70% of the sera. There was a statistically significant association between the detection of B19 DNA by PCR and high B19 IgM values (P < 0.005), low B19 IgG values (P < 0.05), and a short interval between onset of symptoms and serum collection (P < 0.005). Serial serum samples, throat swabs, and peripheral blood mononuclear cells collected from 10 individuals during an outbreak of parvovirus B19 were also tested by the nested PCR. B19 DNA was detectable in the throat swabs at the time of the clinical illness and in the peripheral blood mononuclear cell fraction up to the end point of the study 6 months after infection. The location of the B19 DNA could not be determined in cytocentrifuge preparations of peripheral blood mononuclear cells with nonisotopic in situ hybridization and immunolabelling.
本文描述了一种用于检测人细小病毒B19 DNA的巢式聚合酶链反应（PCR）检测方法的特性及应用。该检测方法用149份诊断血清样本进行了评估（这些样本在症状出现后150天内采集），这些样本之前已通过斑点杂交检测B19 DNA，并通过类别特异性捕获放射免疫测定法检测B19免疫球蛋白M（IgM）和IgG。PCR可在70%的血清中检测到B19 DNA。PCR检测到的B19 DNA与高B19 IgM值（P < 0.005）、低B19 IgG值（P < 0.05）以及症状出现与血清采集之间的短间隔（P < 0.005）之间存在统计学上的显著关联。在细小病毒B19爆发期间从10名个体采集的系列血清样本、咽拭子和外周血单个核细胞也通过巢式PCR进行了检测。在临床疾病发作时，咽拭子中可检测到B19 DNA，在感染后6个月研究结束时，外周血单个核细胞部分中也可检测到B19 DNA。在外周血单个核细胞的细胞离心涂片制备物中，无法通过非同位素原位杂交和免疫标记确定B19 DNA的位置。
