【作者】 江凌晓；

【导师】 曹虹；

【作者基本信息】 第一军医大学 ， 病原生物学， 2000， 硕士

【摘要】 人微小病毒B19（human parvovirus B19，PVB19）是微小病毒科微小病毒属中目前已知的唯一致人类疾病的病毒，也是动物病毒中体积最小结构最简单的DNA病毒。现已证实该病毒同多种儿童及成人疾病有关，且随着检测手段的改进及研究的深入，其疾病谱还在不断扩大。 本研究应用巢式聚合酶链反应（Nest-PCR）对473例临床标本进行了检测，共发现17例阳性标本。其阳性疾病谱分别为过敏性紫癜1例，急性淋巴细胞白血病2例，急性非淋巴细胞白血病1例，再生障碍性贫血1例，肝癌术后1例，乙型病毒性肝炎4例，丙型病毒性肝炎1例，非甲-戊型肝炎2例，查体（无症状）2例，皮疹1例，脑炎1例。 17例阳性标本中有2例的临床资料完整而清晰，因此，为初步了解PVB19病毒广东分离株的基因变异情况，我们用加强聚合酶链反应（re-PCR）扩增了这2例阳性标本（一例为活动性肝硬化伴再生障碍性贫血患者，另一例为患急性淋巴细胞白血病的儿童）的PVB19病毒结构蛋白VP1/VP2区（2909-3806 nt）长约898bp的基因片段。该目的片段经TA克隆后进行核苷酸序列测定。测定序列与标准序列比较后发现：同一时间，在广东地区分离的PVB19病毒株核苷酸变异率有很大差别，两株病毒在该区段的核苷酸变异数分别为32和10，对应的氨基酸变异数分别为6和1。通过与Genbank中已有的该区段核苷酸序列进行种系发生树分析后发现PVB19病毒中国流行株属种系发生树分型的Ⅰ型且与欧美、日本、韩国流行株在进化速度上有很大差异。另外还发现PVB19病毒株种系发生树分型结果与感染PVB19后的临床表现关系不明确。更多还原

【Abstract】 Human parvovirus B19 (PVB 19), the only parvovirus known to be pathogenic in human , belongs to the genus Parvovirus in the family Parvoviridae. It is the smallest animal DNA virus. PVB 19 infection often results in a variety of clinical manifestations in adults and children . The objective of this research was to preliminary study the variation of PVB19 in Guangdong Province. Firstly, we used Nest-PCR method to detect 496 serum samples obtained from 473 patients. 18 sampels obtained from 17 patients were positive for PVB 19 DNA. Of the 17 patients, two had clear clinical record. one was a child with acute lymphocytic leukemia, the other was a adult with chronic active hepatitis B associated with aplastic anemia .Then a methodology combined with re-PCR amplifying, TA cloning and nucleotide sequencing was used to analyses the 898bp fragment (2909-3806 nt) of the two strains Compared with standard strain, the rate of nucleotide variability of the two strains was very different, even the. two strains were isolated in Guangdong Province in the same time. We also compared the two strain抯 fragmentes with other associated strains抯 fragment, which was searched from Genbank . These data suggest that PVB 19 isolates from China were significantly different from other isolates at both the nucleotide and prediected amino acid levels, and that no particular genotype of PVB19 was associated with the particular clinical outcome.更多还原