应用PCR法对临床血标本中人微小病毒B19的检测分析

Polymerase Chain Reaction Assay of Human Parvovirus B19 in Clinical Blood Specimens

目的 探讨 PCR法检测临床血标本中人微小病毒 B1 9( HPV B1 9)的应用价值。方法 根据序列比对结果 ,在HPV B1 9核苷酸相对保守区设计引物进行 PCR扩增。应用双脱氧链末端终止法对 HPV B1 9阳性 PCR产物进行克隆测序。结果 应用该法最终检出HPV B1 9的血清稀释度为 1 0 3。序列比较表明 ,用本法检出的 1株HPV B1 9阳性标本与标准株 ( Au株 )核苷酸序列同源性为 92 %。检测肝癌和肝硬化患者血清标本各 1 4例 ,结果 HPV B1 9阳性分别为 8例和 5例。结论 本法可用于 HPV B1

Objective To detect human parvovirus B19 in clinical blood samples by polymerase chain reaction (PCR). Methods The primers was derived from the conserved region of HPV B19 according to the result of sequence comparing. The product of the PCR was cloned into plasmid and sequenced by the dideoxy-mediated chain-termination method. Results The maximum dilution of serum was 10 3 for detection of HPV B19 by PCR. Sequence analysis showed nucleotide homology was 92% between the cloned HPV B19 and the standard strain (Au strain) registered in GenBank. The rate of HPV B19 infections were 8/14 and 5/14 in patients with hepatocellular carcinoma and hepatocirrhosis, respectively. Conclusion This method may be used for the clinical diagnosis and epidemiological study of HPV B19 infection.